Single-cell dissection of a rare human prostate …specimens collected, after dissociation of the...

1

Single-cell dissection of a rare human prostate basal cell carcinoma

Summary heading：Single-cell analysis of prostate basal cell carcinoma

Abstract

As a rare subtype of prostate carcinoma, basal cell carcinoma (BCC) has not been

studied extensively and thus lacks systematic molecular characterization. Here we

applied single-cell genomic amplification and RNA-Seq to a specimen of human

prostate BCC (CK34βE12+/P63+/PAP-/PSA-). The mutational landscape was obtained

via whole exome sequencing of the amplification mixture of 49 single cells, and the 5

putative driver genes mutated are CASC5, NUTM1, PTPRC, KMT2C and TBX3. The

top 3 nucleotide substitutions are C>T, T>C and C>A, similar to common prostate

cancer. The distribution of the variant allele frequency values indicated these single

cells are from the same tumor clone. The transcriptomes of 69 single cells were

obtained, and they were clustered into tumor, stromal and immune cells based on their

global transcriptomic profiles. The tumor cells specifically express basal cell markers

like KRT5, KRT14 and KRT23, and epithelial markers EPCAM, CDH1 and CD24.

The transcription factor (TF) co-variance network analysis showed that the BCC

tumor cells have distinct regulatory networks. By comparison with current prostate

cancer datasets, we found that some of the bulk samples exhibit basal-cell signatures.

Interestingly, at single-cell resolution the gene expression patterns of prostate BCC

tumor cells show uniqueness compared with that of common prostate cancer-derived

circulating tumor cells. This study, for the first time, discloses the comprehensive

mutational and transcriptomic landscapes of prostate BCC, which lays a foundation

certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. The copyright holder for this preprint (which was notthis version posted October 25, 2019. . https://doi.org/10.1101/818260doi: bioRxiv preprint

https://doi.org/10.1101/818260

2

for the understanding of its tumorigenesis mechanism and provides new insights into

prostate cancers in general.

Xianbin Su1,2, #, Qi Long3, #, Juanjie Bo1, #, Yi Shi2, #, Li-Nan Zhao2, Yingxin Lin4,

Qing Luo2, Shila Ghazanfar4,5, Chao Zhang6, Qiang Liu7, Lan Wang2, Kun-Yan He2,

Jian He2, Xiao-Fang Cui2, Jean Y. H. Yang4, Ze-Guang Han2,*, Jian-Jun Sha1,*,

Guoliang Yang1, *

1Department of Urology and Key Laboratory of Systems Biomedicine (Ministry of

Education), Renji Hospital, School of Medicine, Shanghai Jiao Tong University,

Shanghai, China

2Key Laboratory of Systems Biomedicine (Ministry of Education) and Collaborative

Innovation Center of Systems Biomedicine, Shanghai Center for Systems

Biomedicine, Shanghai Jiao Tong University, Shanghai, China

3Key Laboratory for Regenerative Medicine (Ministry of Education), School of

Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong,

Hong Kong SAR, China

4Charles Perkins Center and School of Mathematics and Statistics, The University of

Sydney, Sydney, Australia

5Cancer Research UK Cambridge Institute, University of Cambridge, Li Ka Shing

Centre, Robinson Way, Cambridge CB2 0RE, United Kingdom

6Department of Urology, Shanghai Changhai Hospital, Second Military Medical


https://doi.org/10.1101/818260

3

University, Shanghai, China

7Department of Pathology, Renji Hospital, School of Medicine, Shanghai Jiao Tong

University, Shanghai, China

#These authors contributed equally.

*To whom correspondence should be addressed:

Guoliang Yang, Department of Urology, Renji Hospital, School of Medicine,

Shanghai Jiao Tong University, Shanghai 200127, China. Tel: +86-21-6373 0455; Fax:

+86-21-6373 0455; Email: [email protected]

Jian-Jun Sha, Department of Urology, Renji Hospital, School of Medicine, Shanghai

Jiao Tong University, Shanghai 200127, China. Tel: +86-21-6373 0455; Fax:

+86-21-6373 0455; Email: [email protected]

Ze-Guang Han, Key Laboratory of Systems Biomedicine (Ministry of Education),

Shanghai Center of Systems Biomedicine, Shanghai Jiao Tong University, Shanghai

200240, China. Tel: +86-21-3420 7304; Fax: +86-21-3420 6059; Email:

[email protected]


https://doi.org/10.1101/818260

4

Running title

Single-cell analysis of prostate basal cell carcinoma

Abbreviations

BCC, basal cell carcinoma; CTCs, circulating tumor cells; MDA, multiple

displacement amplification; PAP, prostatic acid phosphatase; PSA, prostate-specific

antigen; TF, transcription factor; VAF, variant allele frequency.

Keywords

prostate, basal cell carcinoma, single-cell sequencing, gene expression, transcription

factor, mutation

Conflict of Interest

The authors declare that they have no conflicts of interest with the contents of this

article.


https://doi.org/10.1101/818260

5

Introduction

Prostate cancer is a heterogeneous disease with complex subtypes and different cell

origins [1-3], and basal cell carcinoma (BCC) is an extremely rare histological

subtype comprising < 0.01% of prostate cancer [4]. Despite being largely regarded as

pursuing an indolent clinical course, aggressive behaviors like recurrence or

metastasis have been observed and deaths also reported, highlighting the complex and

yet poorly understood mechanism [5-9]. There have been extensive reports on the

genomic features of common prostate cancer [10-17], and the molecular features of

normal human prostate basal cells, prostate basal cell hyperplasia, and basal

populations from human prostate cancer have also been reported [18-20], but the

molecular profile of prostate BCC is still lacking. The mutational and transcriptomic

features of this tumor sub-type will thus be valuable for understanding of its

tumorigenesis mechanism and development of future clinical treatments.

Single-cell sequencing has evolved to be a powerful tool that provides

unprecedented resolution of clinical specimens especially with limited amounts, and

marker-free decomposition of the constituent cell types based on single-cell

transcriptional profiles allows precise dissection of complex systems such as various

tissues or tumors [21-23]. Here we applied single-cell genomic amplification to a

human specimen of prostate BCC and used the mixture of 49 single cells to provide

mutational landscape of prostate BCC via whole exome sequencing. We then used

single-cell RNA-Seq to obtain the transcriptomes of 69 cells from the same specimen.

Three types of cells, namely tumor, stromal and immune cells, were identified with


https://doi.org/10.1101/818260

6

their molecular features revealed at single-cell resolution, providing a useful resource

for not only this prostate cancer subtype but prostate cancer in general.

Results

Histological and immunohistochemical features of human prostate BCC

The tumor exhibited a widespread infiltrative growth pattern under microscopic

examination, and individual cells had large pleomorphic nuclei and scant cytoplasm

(Fig. 1). The tumor was negative for two known prostatic markers, prostatic acid

phosphatase (PAP) and prostate-specific antigen (PSA), but strongly expressed

CK34βE12 and P63, suggestive of basal cell origin (Fig. 1). Based on the cellular

morphology and immunohistochemical staining, we diagnosed it as prostate BCC.

The patient with BCC was resistant to chemotherapy but sensitive to radiotherapy,

and followed-up for 32 months without recurrence or metastasis (Supporting

Information, Fig. S1).

Whole exome sequencing reveals the mutational landscape of prostate BCC

As genetic variations are generally believed to be the causes of tumors, here we tried

to reveal the mutational features of prostate BCC. Due to the limited amount of tumor

specimens collected, after dissociation of the specimens into single-cell suspension,

we first conducted single-cell whole genome multiple displacement amplification

(MDA) on a microfluidic-chip. We then used the mixture of amplified products from

49 single cells for whole exome sequencing (Fig. 2A). A total of 91 functional exonic

mutations were obtained, and 5 of the mutated genes, CASC5, NUTM1, PTPRC,


https://doi.org/10.1101/818260

7

KMT2C and TBX3 are putative driver genes catalogued in the COSMIC Cancer Gene

Census (Supporting Information, Fig. S2) [24]. Mutations in DNA repair genes are

related to poor outcomes of prostate cancers [12, 25], but few are mutated in this

prostate BCC sample such as BRCA2, consistent with good survival of this patient.

The top 3 nucleotide substitutions are C>T, T>C and C>A, which are exactly the

same top 3 substitutions in recently reported localized, non-indolent prostate tumors

(Fig. 2B) [13]. This suggests the etiology of prostate BCC is likely the same as

common prostate tumors. The major composite COSMIC signatures are Signature 4,

5, 6 and 12 (Fig. 2C). While the etiologies of Signature 5 and 12 are still unknown,

Signature 4 and 6 are associated with smoking and defective DNA mismatch repair,

respectively [26].

By checking the distribution of the variant allele frequency (VAF) of the exonic

mutations of prostate BCC, it is clear that most of the variations have VAF values of

around 0.5, thus supporting that most of the single cells share a similar mutational

background (Fig. 2D). The VAF values of the 5 driver mutations are also

approximately 0.5 (Fig. 2E), indicating these single cells are probably originated from

the same ancestor and form the major tumor clone. However, there are still some

variations with lower VAF values which suggest sub-clonal mutations may have

existed in some cells that are acquired during later diversified evolution of the tumor.

To understand whether the prostate BCC is showing similarities to other reported

prostate cancer cases, we checked the mutational frequency of the prostate BCC

mutated genes in prostate cancer samples collected in cBioPortal [27]. For the 88


https://doi.org/10.1101/818260

8

genes enquired, only 10 genes are not mutated, and 27 genes are mutated in more than

0.5% of the ~4400 prostate cancer samples (Fig. 2F). Interestingly, 3 of the driver

genes from this study, KMT2C, PTPRC, and TBX3 are also among the top mutated

genes, especially KMT2C with mutation rate of ~6%. GSEA/MSigDB enrichment

analysis [28] of the mutated genes shows that the top enriched terms are related to cell

cycle, cytoskeleton and others (Fig. 2G), consistent with its basal cell origin.

Single-cell RNA-Seq reveals the transcriptomic features of prostate BCC

To understand the transcriptional profiles of prostate BCC, we then randomly selected

69 single cells for single-cell RNA-Seq (Fig. 3A). With a median unique mapping rate

of 58.4% and 1.08 million mapped reads, we obtained a median of ~2800 genes per

cell (Supporting Information, Fig. S3A). We used ERCC spike-ins as positive controls,

and the high correlation efficiency between single-cells based on the 92 spike-ins

confirmed the high quality of the data (Supporting Information, Fig. S3B-D). After

filtering 5 outliers, the remaining 64 cells were classified into three clusters based on

their global transcriptomic profiles (Fig. 3B-C). GSEA/MSigDB enrichment analysis

identified them as tumor cells (51 cells), stromal cells (7 cells) and immune cells (6

cells) (Fig. 3D).

The three clusters of cells can also be clearly separated in tSNE plot based on the

top genes expressed in each cluster (Fig. 4A). The tumor cells specifically expressed

genes encoding cytokeratins including KRT5, KRT14 and KRT23, which are reliable

basal cell markers (Fig. 4B). Interestingly, epithelial markers EPCAM, CDH1 and


https://doi.org/10.1101/818260

9

CD24 are also highly expressed in tumor cells. Stromal cells specifically express

COL1A2, THY1 and BGN, and immune cells specifically express monocytic marker

CD14, CXCL8 and myeloid cell nuclear differentiation antigen MNDA (Fig. 4B),

suggestive of possible innate immune cell infiltration. Immunohistochemical staining

confirmed the expression of CK14 (KRT14) at protein level, further proving the

reliability of the sequencing data (Fig. 4C).

Transcription factors (TFs) covariance network analysis showed that the three

clusters demonstrate different transcriptional regulatory status (Fig. 4D). Significantly,

MYC, ARID1A, TBX3 and SOX11 are highly expressed in tumor cells, where, besides

the well-known MYC that is recently reported to be over-expressed in human prostatic

basal cells [19], the oncogenic role of ARID1A, a key component of chromatin

remodeling complex SWI/SNF, has been recently demonstrated in liver cancer [29],

the development-related gene SOX11 was reported to be elevated in basal-like breast

cancer [30], and TBX3 plays important roles in development, differentiation and

tumorigenesis [31]. The TFs co-variance network provides clues to future dissection

of the regulatory mechanism of BCC.

Single-cell analysis of prostate BCC provides new insights to prostate cancer

We then checked the expression of prostate BCC tumor cell specific genes in

previously reported luminal and basal cells sorted from normal and cancerous

prostates by flow cytometry [18]. The prostate basal cells exhibit higher expression

levels of KRT5, KRT14 and KRT23 compared with luminal cells, similarly as our


https://doi.org/10.1101/818260

10

prostate BCC tumor cells, further confirming their basal cell origin (Fig. 5A).

Interestingly, the genes highly expressed in prostate BCC tumor cells fall into two

groups based on their expression correlation patterns, with one group including KRT5,

KRT14, KRT23, DSC3, FGFR2, etc., while the other group including EPCAM, CDH1,

CD24, etc. (Fig. 5A). It seems the former group includes the basal cell markers while

the latter group includes genes expressed in common prostate epithelial cells.

We further checked the expression patterns of BCC tumor cell specific genes in

other bulk samples of human prostate cancers [10, 11, 32]. It is interesting to identify

a small group of samples exhibiting basal cell gene expression features in two

separate prostate cancer datasets (shaded branches in the dendrograms of Fig. 5B and

Supporting Information, Fig. S4A). Likely explanations are that these tumor samples

were derived from BCC patients or the samples contained basal cells. For TCGA

PRAD dataset, the expressions of basal markers are detected in most samples,

possibility due to the high number of genes detected by deep sequencing (Supporting

Information, Fig. S4B). The split of the genes highly expressed in BCC tumor cells

into 2 groups is further confirmed by these three datasets, with KRT5, KRT14, KRT23

clustering together with DSC3, FGFR2 and also possibly ACTG2. Our single-cell

analysis of prostate BCC reveals the existence of basal cell features in current bulk

samples of prostate cancer.

We then compared our results with one single-cell resolution dataset of prostate

cancer [33]. Compared with circulating tumor cells (CTCs) from prostate cancer

patients, the prostate BCC tumor cells exhibit distinct gene expression features with


https://doi.org/10.1101/818260

11

high levels of IGF2, KRT5, KRT14, KRT23, AQP1, MIA, FGFR2 and ACTG2,

although they all expressed EPCAM, CDH1 and CD24 (Fig. 5C). The results

demonstrated the unique gene expression patterns of the prostate BCC tumor cells,

which exhibit the molecular features of basal cells with activated genes like IGF2 and

FGFR2, consistent with report on the regulation of pluripotency by IGF and FGF

pathways [34]. Another interesting finding is that the BCC immune cells clustered

together with some single cells annotated as CTC-candidate, which were likely to be

contaminating leukocytes to the CTCs (Fig. 5C). The single-cell level data also

support the above mentioned split of the genes highly expressed in prostate BCC

tumor cells, with one group representing genuine BCC markers while the other genes

commonly expressed in prostate cancer.

Detection of exonic mutations in single-cell RNA-Seq reads

As our mutational data and single-cell RNA-Seq data were generated from the same

prostate BCC specimen, we checked whether the exonic mutations could be detected

in RNA-Seq reads. Interestingly, we indeed found mutations in RNA-Seq reads from

some single cells for GABPB2, ASTE1, COPS3 and BEX2 (Supporting Information,

Fig. S5). The variations are only detected in some of the single cells, which are likely

because the mutated sites are not always covered by the relatively shallow single-cell

RNA-Seq. Another phenomenon is that single cells in RNA-Seq analysis only

contained either reference or altered variant, which is probably caused by allelic


https://doi.org/10.1101/818260

12

specific expression or allele dropout in RNA-Seq. The consistency of scRNA-Seq

data and exome-seq data further suggested the reliability of our prostate BCC data.

Discussion

As a rare malignancy, there has been little research into the molecular characteristics

of prostate BCC and no consensus on its treatment. Mostly, such data exist as

sporadic reports on immunohistochemical expression in a small number of cases [8,

35-37]. This study represents the first genomic and transcriptomic profiling of this

prostate cancer subtype, and the single-cell resolution transcriptomic data is especially

valuable for understanding of not only this subtype but prostate cancer in general.

The molecular features of normal human prostate basal cells [19], prostate basal

cell hyperplasia [20], and human prostate cancer derived basal populations [18] have

been reported recently, and the single-cell transcriptomic profiles of prostate BCC is a

good complement to these data for comparison studies and better understanding of

prostate cancer cell origin. Most of the current genomics data for prostate cancer are

generated using bulk samples [10, 11, 13], and the single-cell transcriptional profiles

of prostate BCC provide an opportunity for deeper utilization of these data. For

example, analysis using genes highly expressed in prostate BCC reveals the existence

of basal cell features in some bulk samples of prostate cancer, which is consistent

with recent classification of prostate cancer samples into basal-like and luminal-like

subtypes using the PAM50 classifier [1, 38]. The single-cell resolution profiles of

constitutional cell types from prostate BCC provide further advantage for future


https://doi.org/10.1101/818260

13

de-convolution of bulk samples [39], which will maximize the values of current

genomic data of prostate cancer.

The prostate BCC patient is still surviving now, indicating this case is an indolent

malignancy. Our genomics and transcriptomic analysis of this chemotherapy-resistant

but radiotherapy-sensitive prostate BCC provides a useful resource for future

dissection of the molecular mechanisms related to the radiotherapy responsiveness

and treatment guidance. Mutations that disrupt the function of DNA damage repair

genes have been shown to be associated with the aggressive clinical behavior of

localized prostate cancer and with cancer-specific mortality [12, 40-42], and it has

also been reported that men with metastatic prostate cancer and DNA-repair gene

mutations have sustained responses to poly-ADP ribose polymerase (PARP)

inhibitors and platinum-based chemotherapy [43, 44]. This study showed that the

prostate BCC patient with few DNA-repair gene mutations was resistant to

chemotherapy, suggesting that we may directly adopt radiotherapy rather than

chemotherapy for such conditions. The results indicated that precision treatment of

BCC could be more than just a histopathological triviality but be based on association

of tumor phenotypes with molecular features.

In summary, single-cell analysis revealed the genomic and transcriptomic

landscapes of the rare prostate BCC for the first time. This provides clues for

elucidation of the tumorigenesis mechanism, further discovery of biomarkers and

therapeutic targets, and better understanding of not only prostate BCC but prostate

cancer in general.


https://doi.org/10.1101/818260

14

Methods

Clinical specimens

This study was approved by the Ethnical Review Board of Renji Hospital, School of

Medicine, Shanghai Jiao Tong University. Specimens were obtained by prostatic

needle biopsy from a 55-year-old man with pelvic pain and irritative urinary

symptoms. The specimens underwent pathological evaluation. After diagnosis as

prostate BCC, the patient was treated with chemotherapy and radiotherapy and then

followed-up for 32 months without evidence of local recurrence or distant metastases

(Supporting Information, Fig. S1). The specimens were mechanically dissociated and

digested with collagenase IV and DNase I, and the single-cell suspensions were used

for both single-cell genome amplification and single-cell RNA-Seq.

Immunohistochemical staining

Immunohistochemical staining was done as previously described [45], and we used

the following primary antibodies: P63 (abcam, ab124762), CK34βE12 (Dako,

M0630), CK14 (abcam, ab7800), PAP (Dako, M0792), PSA (Dako, A056201). We

used biotinylated universal link antibody as the secondary antibody.

Single-cell genome amplification and whole exome sequencing

Single-cell capture, lysis, reverse-transcription, and whole genome MDA were done

in a microfluidic-based C1 DNA-Seq IFC (10~17 μm, Fluidigm) according to its


https://doi.org/10.1101/818260

15

protocol using illustra GenomiPhi V2 DNA Amplification Kit (GE Healthcare,

25660031). The amplified products from 49 single cells were mixed for exonic region

capture and library preparation using Agilent SureSelect Human All Exon v7 Kit

(Agilent, 5191-4005). The library was sequenced using illumina HiSeq platform with

2 × 150 bp sequencing mode. We used GATK [46] for variant calling and ANNOVAR

[47] for functional annotation of the mutations. As there is no para-tumor tissue for

this patient, we filtered SNPs using dbSNP141 [48] and 1,000 Genomes Project (v3)

database [49]. We used MutationalPatterns [50] to decipher the mutational signature

composition. For genes mutated in this prostate BCC sample, we checked their

mutational frequencies in prostate cancer samples collected in cBioPortal [27].

GSEA/MSigDB enrichment analysis was also conducted for the genes mutated in this

prostate BCC.

Single-cell RNA-Seq and data analysis

Single-cell RNA-Seq experiment and analysis were conducted as previously

described [45]. Single-cell capture, lysis, reverse-transcription, and cDNA

amplification were done in a microfluidic-based C1 RNA-Seq IFC (10~17 μm,

Fluidigm). After library preparation with Nextera XT Kit and Index Kit (illumina), the

single-cell libraries were pooled and sequenced using NextSeq 500 (illumina) with 2

× 76 bp sequencing mode. After generation of FPKM data, SC3 [51] was used for

identification of cell outliers and single-cell clustering. The genes differently

expressed between clusters were then identified via ANOVA analysis (p < 0.05) using


https://doi.org/10.1101/818260

16

SINGuLAR™ (Fluidigm). GSEA/MSigDB enrichment analysis was conducted for

the genes specifically expressed in each cluster to facilitate cell type identification.

Transcription factors (TFs) covariance network analysis was conducted to reveal the

relationship of the TFs specifically expressed in each cell cluster as previously

described [45, 52]. Gene expression files were retrieved from five other prostate

cancer studies [10, 11, 18, 32, 33] for comparison studies.

Acknowledgement

This work is supported in part by the National Natural Science Foundation of China

(81802806, 81472621, 81402329 and 81902561), National Program on Key

Research Project of China (2016YFC0902701, Precision Medicine), Shanghai Health

Bureau (20164Y0124), Medical and Engineering Crossover Fund of SJTU

(YG2016QN71, YG2017MS67), University of Sydney – Shanghai Jiao Tong

University Joint Research Alliance (USYD-SJTU JRA) grant, funding from Key

Laboratory of Systems Biomedicine (Ministry of Education) (KLSB2017QN-03), and

Incubating Program for Clinical Research and Innovation of Renji Hospital SJTU

School of Medicine (PYII-17-010).

Authors' contributions

XS and GY designed the study. XS, JB, LZ, CZ, QL3, LW, KH, JH, XC, WX and GY

carried out experiments. XS, QL1, YS, YL, QL2, SG and JY analyzed data. XS, GY

and ZH drafted the manuscript. GY, JS and ZH supervised the study. All authors read

and approved final version of the manuscript.


https://doi.org/10.1101/818260

17

References

1. Zhao SG, Chang SL, Erho N, Yu M, Lehrer J, Alshalalfa M, Speers C,

Cooperberg MR, Kim W, Ryan CJ, et al. (2017) Associations of Luminal and Basal

Subtyping of Prostate Cancer With Prognosis and Response to Androgen Deprivation

Therapy. JAMA Oncol 3, 1663-1672.

2. Goldstein AS, Huang J, Guo C, Garraway IP & Witte ON (2010) Identification of

a cell of origin for human prostate cancer. Science 329, 568-571.

3. Choi N, Zhang B, Zhang L, Ittmann M & Xin L (2012) Adult murine prostate

basal and luminal cells are self-sustained lineages that can both serve as targets for

prostate cancer initiation. Cancer Cell 21, 253-265.

4. Tannenbaum M (1975) Adenoid cystic or "salivary gland" carcinomas of prostate.

Urology 6, 238-239.

5. Halat SK & MacLennan GT (2008) Adenoid cystic/basal cell carcinoma of the

prostate. J Urol 179, 1576.

6. Komura K, Inamoto T, Tsuji M, Ibuki N, Koyama K, Ubai T, Azuma H &

Katsuoka Y (2010) Basal cell carcinoma of the prostate: unusual subtype of prostatic

carcinoma. Int J Clin Oncol 15, 594-600.

7. Chang K, Dai B, Kong Y, Qu Y, Wu J, Ye D, Yao X, Zhang S, Zhang H, Zhu Y, et

al. (2013) Basal cell carcinoma of the prostate: clinicopathologic analysis of three

cases and a review of the literature. World J Surg Oncol 11, 193.

8. Ali TZ & Epstein JI (2007) Basal cell carcinoma of the prostate: a

clinicopathologic study of 29 cases. Am J Surg Pathol 31, 697-705.

9. Ayyathurai R, Civantos F, Soloway MS & Manoharan M (2007) Basal cell

carcinoma of the prostate: current concepts. BJU Int 99, 1345-1349.

10. The Cancer Genome Atlas Research Network (2015) The Molecular Taxonomy


https://doi.org/10.1101/818260

18

of Primary Prostate Cancer. Cell 163, 1011-1025.

11. Robinson D, Van Allen EM, Wu YM, Schultz N, Lonigro RJ, Mosquera JM,

Montgomery B, Taplin ME, Pritchard CC, Attard G, et al. (2015) Integrative clinical

genomics of advanced prostate cancer. Cell 161, 1215-1228.

12. Pritchard CC, Mateo J, Walsh MF, De Sarkar N, Abida W, Beltran H, Garofalo A,

Gulati R, Carreira S, Eeles R, et al. (2016) Inherited DNA-Repair Gene Mutations in

Men with Metastatic Prostate Cancer. N Engl J Med 375, 443-453.

13. Fraser M, Sabelnykova VY, Yamaguchi TN, Heisler LE, Livingstone J, Huang V,

Shiah YJ, Yousif F, Lin X, Masella AP, et al. (2017) Genomic hallmarks of localized,

non-indolent prostate cancer. Nature 541, 359-364.

14. Grasso CS, Wu YM, Robinson DR, Cao X, Dhanasekaran SM, Khan AP, Quist

MJ, Jing X, Lonigro RJ, Brenner JC, et al. (2012) The mutational landscape of lethal

castration-resistant prostate cancer. Nature 487, 239-243.

15. Quigley DA, Dang HX, Zhao SG, Lloyd P, Aggarwal R, Alumkal JJ, Foye A,

Kothari V, Perry MD, Bailey AM, et al. (2018) Genomic Hallmarks and Structural

Variation in Metastatic Prostate Cancer. Cell 174, 758-769.

16. Espiritu SMG, Liu LY, Rubanova Y, Bhandari V, Holgersen EM, Szyca LM, Fox

NS, Chua MLK, Yamaguchi TN, Heisler LE, et al. (2018) The Evolutionary

Landscape of Localized Prostate Cancers Drives Clinical Aggression. Cell 173,

1003-1013 e1015.

17. Baca SC, Prandi D, Lawrence MS, Mosquera JM, Romanel A, Drier Y, Park K,

Kitabayashi N, MacDonald TY, Ghandi M, et al. (2013) Punctuated evolution of

prostate cancer genomes. Cell 153, 666-677.

18. Smith BA, Sokolov A, Uzunangelov V, Baertsch R, Newton Y, Graim K, Mathis

C, Cheng D, Stuart JM & Witte ON (2015) A basal stem cell signature identifies


https://doi.org/10.1101/818260

19

aggressive prostate cancer phenotypes. Proc Natl Acad Sci U S A 112, E6544-6552.

19. Zhang D, Park D, Zhong Y, Lu Y, Rycaj K, Gong S, Chen X, Liu X, Chao HP,

Whitney P, et al. (2016) Stem cell and neurogenic gene-expression profiles link

prostate basal cells to aggressive prostate cancer. Nat Commun 7, 10798.

20. Henry G, Malewska A, Mauck R, Gahan J, Hutchinson R, Torrealba J, Francis F,

Roehrborn C & Strand D (2017) Molecular pathogenesis of human prostate basal cell

hyperplasia. Prostate 77, 1344-1355.

21. Gao S, Yan L, Wang R, Li J, Yong J, Zhou X, Wei Y, Wu X, Wang X, Fan X, et al.

(2018) Tracing the temporal-spatial transcriptome landscapes of the human fetal

digestive tract using single-cell RNA-sequencing. Nat Cell Biol 20, 721-734.

22. Han X, Wang R, Zhou Y, Fei L, Sun H, Lai S, Saadatpour A, Zhou Z, Chen H, Ye

F, et al. (2018) Mapping the Mouse Cell Atlas by Microwell-Seq. Cell 172, 1091-1107

e1017.

23. Tanay A & Regev A (2017) Scaling single-cell genomics from phenomenology to

mechanism. Nature 541, 331-338.

24. Sondka Z, Bamford S, Cole CG, Ward SA, Dunham I & Forbes SA (2018) The

COSMIC Cancer Gene Census: describing genetic dysfunction across all human

cancers. Nat Rev Cancer 18, 696-705.

25. Taylor RA, Fraser M, Rebello RJ, Boutros PC, Murphy DG, Bristow RG &

Risbridger GP (2019) The influence of BRCA2 mutation on localized prostate cancer.

Nat Rev Urol 16, 281-290.

26. Alexandrov LB, Nik-Zainal S, Wedge DC, Aparicio SA, Behjati S, Biankin AV,

Bignell GR, Bolli N, Borg A, Borresen-Dale AL, et al. (2013) Signatures of

mutational processes in human cancer. Nature 500, 415-421.

27. Gao J, Aksoy BA, Dogrusoz U, Dresdner G, Gross B, Sumer SO, Sun Y,


https://doi.org/10.1101/818260

20

Jacobsen A, Sinha R, Larsson E, et al. (2013) Integrative analysis of complex cancer

genomics and clinical profiles using the cBioPortal. Sci Signal 6, pl1.

28. Subramanian A, Tamayo P, Mootha VK, Mukherjee S, Ebert BL, Gillette MA,

Paulovich A, Pomeroy SL, Golub TR, Lander ES, et al. (2005) Gene set enrichment

analysis: a knowledge-based approach for interpreting genome-wide expression

profiles. Proc Natl Acad Sci U S A 102, 15545-15550.

29. Sun X, Wang SC, Wei Y, Luo X, Jia Y, Li L, Gopal P, Zhu M, Nassour I, Chuang

JC, et al. (2017) Arid1a has context-dependent oncogenic and tumor suppressor

functions in liver cancer. Cancer Cell 32, 574-589 e576.

30. Shepherd JH, Uray IP, Mazumdar A, Tsimelzon A, Savage M, Hilsenbeck SG &

Brown PH (2016) The SOX11 transcription factor is a critical regulator of basal-like

breast cancer growth, invasion, and basal-like gene expression. Oncotarget 7,

13106-13121.

31. Wansleben S, Peres J, Hare S, Goding CR & Prince S (2014) T-box transcription

factors in cancer biology. Biochim Biophys Acta 1846, 380-391.

32. Beltran H, Prandi D, Mosquera JM, Benelli M, Puca L, Cyrta J, Marotz C,

Giannopoulou E, Chakravarthi BV, Varambally S, et al. (2016) Divergent clonal

evolution of castration-resistant neuroendocrine prostate cancer. Nat Med 22,

298-305.

33. Miyamoto DT, Zheng Y, Wittner BS, Lee RJ, Zhu H, Broderick KT, Desai R, Fox

DB, Brannigan BW, Trautwein J, et al. (2015) RNA-Seq of single prostate CTCs

implicates noncanonical Wnt signaling in antiandrogen resistance. Science 349,

1351-1356.

34. Bendall SC, Stewart MH, Menendez P, George D, Vijayaragavan K,

Werbowetski-Ogilvie T, Ramos-Mejia V, Rouleau A, Yang J, Bosse M, et al. (2007)

IGF and FGF cooperatively establish the regulatory stem cell niche of pluripotent


https://doi.org/10.1101/818260

21

human cells in vitro. Nature 448, 1015-1021.

35. Yang XJ, McEntee M & Epstein JI (1998) Distinction of basaloid carcinoma of

the prostate from benign basal cell lesions by using immunohistochemistry for bcl-2

and Ki-67. Hum Pathol 29, 1447-1450.

36. Iczkowski KA, Ferguson KL, Grier DD, Hossain D, Banerjee SS, McNeal JE &

Bostwick DG (2003) Adenoid cystic/basal cell carcinoma of the prostate:

clinicopathologic findings in 19 cases. Am J Surg Pathol 27, 1523-1529.

37. Montironi R, Mazzucchelli R, Stramazzotti D, Scarpelli M, Lopez Beltran A &

Bostwick DG (2005) Basal cell hyperplasia and basal cell carcinoma of the prostate: a

comprehensive review and discussion of a case with c-erbB-2 expression. J Clin

Pathol 58, 290-296.

38. Zhao SG, Chen WS, Das R, Chang SL, Tomlins SA, Chou J, Quigley DA, Dang

HX, Barnard TJ, Mahal BA, et al. (2019) Clinical and Genomic Implications of

Luminal and Basal Subtypes Across Carcinomas. Clin Cancer Res 25, 2450-2457.

39. Wang X, Park J, Susztak K, Zhang NR & Li M (2019) Bulk tissue cell type

deconvolution with multi-subject single-cell expression reference. Nat Commun 10,

380.

40. Leongamornlert D, Saunders E, Dadaev T, Tymrakiewicz M, Goh C,

Jugurnauth-Little S, Kozarewa I, Fenwick K, Assiotis I, Barrowdale D, et al. (2014)

Frequent germline deleterious mutations in DNA repair genes in familial prostate

cancer cases are associated with advanced disease. Br J Cancer 110, 1663-1672.

41. Castro E, Goh C, Olmos D, Saunders E, Leongamornlert D, Tymrakiewicz M,

Mahmud N, Dadaev T, Govindasami K, Guy M, et al. (2013) Germline BRCA

mutations are associated with higher risk of nodal involvement, distant metastasis,

and poor survival outcomes in prostate cancer. J Clin Oncol 31, 1748-1757.


https://doi.org/10.1101/818260

22

42. Gallagher DJ, Gaudet MM, Pal P, Kirchhoff T, Balistreri L, Vora K, Bhatia J,

Stadler Z, Fine SW, Reuter V, et al. (2010) Germline BRCA mutations denote a

clinicopathologic subset of prostate cancer. Clin Cancer Res 16, 2115-2121.

43. Cheng HH, Pritchard CC, Boyd T, Nelson PS & Montgomery B (2016) Biallelic

Inactivation of BRCA2 in Platinum-sensitive Metastatic Castration-resistant Prostate

Cancer. Eur Urol 69, 992-995.

44. Mateo J, Carreira S, Sandhu S, Miranda S, Mossop H, Perez-Lopez R, Nava

Rodrigues D, Robinson D, Omlin A, Tunariu N, et al. (2015) DNA-Repair Defects

and Olaparib in Metastatic Prostate Cancer. N Engl J Med 373, 1697-1708.

45. Su X, Shi Y, Zou X, Lu Z-N, Xie G, Yang JYH, Wu C-C, Cui X-F, He K-Y, Luo

Q, et al. (2017) Single-cell RNA-Seq analysis reveals dynamic trajectories during

mouse liver development. BMC Genomics 18, 946.

46. McKenna A, Hanna M, Banks E, Sivachenko A, Cibulskis K, Kernytsky A,

Garimella K, Altshuler D, Gabriel S, Daly M, et al. (2010) The Genome Analysis

Toolkit: a MapReduce framework for analyzing next-generation DNA sequencing

data. Genome Res 20, 1297-1303.

47. Wang K, Li M & Hakonarson H (2010) ANNOVAR: functional annotation of

genetic variants from high-throughput sequencing data. Nucleic Acids Res 38, e164.

48. Sherry ST, Ward MH, Kholodov M, Baker J, Phan L, Smigielski EM & Sirotkin

K (2001) dbSNP: the NCBI database of genetic variation. Nucleic Acids Res 29,

308-311.

49. Auton A, Brooks LD, Durbin RM, Garrison EP, Kang HM, Korbel JO, Marchini

JL, McCarthy S, McVean GA & Abecasis GR (2015) A global reference for human

genetic variation. Nature 526, 68-74.

50. Blokzijl F, Janssen R, van Boxtel R & Cuppen E (2018) MutationalPatterns:


https://doi.org/10.1101/818260

23

comprehensive genome-wide analysis of mutational processes. Genome Med 10, 33.

51. Kiselev VY, Kirschner K, Schaub MT, Andrews T, Yiu A, Chandra T, Natarajan

KN, Reik W, Barahona M, Green AR, et al. (2017) SC3: consensus clustering of

single-cell RNA-seq data. Nat Methods 14, 483-486.

52. Zhang HM, Liu T, Liu CJ, Song S, Zhang X, Liu W, Jia H, Xue Y & Guo AY

(2015) AnimalTFDB 2.0: a resource for expression, prediction and functional study of

animal transcription factors. Nucleic Acids Res 43, D76-81.


https://doi.org/10.1101/818260

24

Figure Legend

Fig. 1. Histological and immunohistochemical staining of the prostate BCC. H&E

staining and the expressions of CK34βE12, P63, PAP and PSA are shown.

Fig. 2. Whole exome sequencing reveals the mutational landscape of prostate BCC.

(A) General description of the exome sequencing of mixtures from genomic

amplification products of prostate BCC single cells and numbers of genetic variations.

(B) Mutational spectra of the prostate BCC based on the variants after SNP filter. (C)

Contributions of COSMIC signatures to the prostate BCC mutational spectra. (D)

Distribution of the variant allele frequency (VAF) of the exonic mutations of prostate

BCC. (E) Summary of the driver (overlapped with The Cancer Gene Census) and

putative sub-clonal (VAF lower than 0.35) mutations of prostate BCC. (F) Percent of

samples mutated in the cBioPortal prostate cancer datasets with those above 1%

shown for the exonic mutated genes in prostate BCC. The red stars indicate Census

driver genes. (G) The items enriched with the exonic mutated genes in prostate BCC

via GSEA/MSigDB canonical pathway enrichment analysis.

Fig. 3. Single-cell RNA-Seq reveals the constituent cell types in the prostate BCC. (A)

Experimental workflow. (B) Heat-map showing the single-cell gene-expression

patterns of the three clusters identified. (C) Heat-map showing the Pearson correlation

coefficients between single-cells. (D) The top 10 items enriched with the gene sets

specifically expressed in each cell cluster via GSEA/MSigDB canonical pathway


https://doi.org/10.1101/818260

25

enrichment analysis.

Fig. 4. Single-cell RNA-Seq reveals the transcriptomic features of prostate BCC. (A)

Heat-map showing the expression patterns of top 20 genes specifically and highly

expressed in each cluster, and t-SNE plot of the three clusters. (B) The expression of

selected markers with the median values and the first and third quartiles shown. (C)

Immunohistochemical staining of the specimen with CK14 antibody. (D) TFs

covariance networks of the single-cell data. Each node represents a TF, and each edge

represents correlation between two TFs. The TFs specifically expressed in each

cluster are colored differently.

Fig. 5. Single-cell analysis of prostate BCC provide new insights to prostate cancer.

(A) (left) Heat-map showing the clustering of FACS sorted luminal (CD49f Lo) and

basal (CD49f Hi) cell populations from human benign and cancerous prostates based

on the genes highly expressed in prostate BCC tumor cells; (right) Heat-map showing

the Pearson correlation coefficients between the genes. (Data from Smith et al. PNAS

2015; 112: E6544-E6552). (B) Analysis of prostate cancer samples in similar

approach as (A). (Data from Robinson et al. Cell 2015; 161: 1215–1228). (C)

Comparison of the gene expression profiles of single cells from Cluster 1~3 of

prostate BCC with human prostate CTCs (CTC-candidate and CTC-confirmed), bulk

primary tumors, single cells from prostate cancer cell lines (Pca cell line) and white

blood cells (Data from Miyamoto et al. Science 2015; 349: 1351-1356).


https://doi.org/10.1101/818260

100× 200×

H&E

CK34βE12

P63

PAP

PSA

Fig 1


https://doi.org/10.1101/818260

Variant Allele Frequency

Nu

mb

er

of V

ari

an

t

0.0 0.2 0.4 0.6 0.8 1.0

01

02

03

0

0 2 4 6 8

GO_regulation_of_cell_cycle

GO_cytoskeleton_organization

GO_microtubule_cytoskeleton_org…

GO_microtubule_based_process

GO_microtubule_bundle_formation

GO_cell_cycle

GO_regulation_of_microtubule_ba…

GO_intracellular_signal_transduction

GO_regulation_of_hydrolase_activity

GO_negative_regulation_of_cataly…

kegg_cell_adhesion_molecules_ca…

GO_cell_cycle_process

GO_single_organism_cellular_loca…

-log10(p)

Group Gene #Chr Position Ref Alt AA change VAF

Driv

er

CASC5 15 40903002 A G S86G 0.68

NUTM1 15 34640367 G C V90L 0.63

PTPRC 1 198713275 T A H769Q 0.59

KMT2C 7 151845915 A T I4366N 0.54

TBX3 12 115112422 C G A420P 0.42

Su

b-c

lon

al

KRT18 12 53343099 G A V48M 0.33

TNRC18 7 5360200 G A P2200L 0.33

KRT18 12 53343132 G T G59W 0.33

SCUBE3 6 35213742 C T T874I 0.30

KRT18 12 53343148 C G A64G 0.27

ART5 11 3661519 T G E47A 0.25

MST1L 1 17084321 A G C566R 0.17

CCDC40 17 78064020 C A T972K 0.12

C7orf26 7 6647724 G C A428P 0.09

0

0.1

0.2

0.3

C>

A

C>

G

C>

T

T>

A

T>

C

T>

G

Pro

port

ion

0

0.2

0.4

0.6

0.8

1

Signature 4

Signature 5

Signature 6

Signature 12

Signature 17

Signature 23

Signature 24

Pro

port

ion

Percent Samples Altered

Whole genome amplification of BCC single cells

Whole exome sequencing (Mixture of 49 cells)

7395 variations after SNP filter

91 exonic variations

5 genes overlap with Census

A B C

D E

F G

Fig 2


https://doi.org/10.1101/818260

Single cells(SC) Captured SC Library prep scRNA-Seq SC transcriptome

0 2 4 6 8 10

reactome_cell_cell_junction_organization

kegg_endometrial_cancer

pid_ajdiss_2pathway

reactome_cell_junction_organization

reactome_cell_cell_communication

kegg_arrhythmogenic_right_ventricular_…

kegg_adherens_junction

kegg_melanoma

kegg_prostate_cancer

kegg_pathways_in_cancer

0 10 20 30 40 50

reactome_axon_guidance

reactome_developmental_biology

reactome_extracellular_matrix_organizati…

pid_avb3_integrin_pathway

naba_matrisome_associated

kegg_focal_adhesion

pid_integrin1_pathway

naba_ecm_glycoproteins

naba_core_matrisome

naba_matrisome

0 10 20 30 40 50

reactome_class_a1_rhodopsin_like_recep…

reactome_g_alpha_i_signalling_events

reactome_cytokine_signaling_in_immune…

reactome_innate_immune_system

kegg_leishmania_infection

reactome_platelet_activation_signaling_a…

kegg_chemokine_signaling_pathway

reactome_hemostasis

reactome_adaptive_immune_system

reactome_immune_system

-log10(p)

Tumor_C02Tumor_C05Tumor_C06Tumor_C07Tumor_C08Tumor_C14Tumor_C15Tumor_C16Tumor_C20Tumor_C22Tumor_C23Tumor_C24Tumor_C27Tumor_C28Tumor_C32Tumor_C33Tumor_C37Tumor_C38Tumor_C40Tumor_C43Tumor_C49Tumor_C50Tumor_C51Tumor_C57Tumor_C58Tumor_C60Tumor_C62Tumor_C63Tumor_C66Tumor_C68Tumor_C70Tumor_C71Tumor_C72Tumor_C73Tumor_C74Tumor_C76Tumor_C77Tumor_C79Tumor_C81Tumor_C82Tumor_C84Tumor_C85Tumor_C86Tumor_C87Tumor_C88Tumor_C89Tumor_C90Tumor_C92Tumor_C93Tumor_C94Tumor_C95Tumor_C01Tumor_C04Tumor_C26Tumor_C41Tumor_C42Tumor_C53Tumor_C69Tumor_C10Tumor_C12Tumor_C19Tumor_C34Tumor_C75Tumor_C91

ZY

XZ

NF

80

4A

ZN

F7

18

ZN

F5

71

ZN

F3

31

ZN

F2

81

ZM

IZ1

ZF

P3

6L

2Z

EB

2-A

S1

ZD

HH

C2

ZB

TB

20

-AS

4Y

WH

AH

YIP

F5

YB

X1

XY

LT

1X

XY

LT

1X

RN

2W

IPF

1W

FD

C2

1P

WD

R4

5B

WA

SH

C4

WA

SH

3P

VS

IG4

VM

O1

VA

V1

VA

SH

1V

AM

P8

UT

P1

4A

US

F2

UR

B1

-AS

1U

CP

2U

BX

N2

BU

BE

2D

1U

BA

C1

TY

RO

BP

TX

ND

C1

1T

TC

38

TS

PA

N3

3T

SP

AN

14

TR

EM

2T

RE

M1

TR

AP

PC

2L

TP

P1

TP

M3

TP

D5

2L

2T

OX

4T

NF

SF

13

BT

NF

SF

13

TN

FR

SF

1B

TN

FA

IP3

TN

FA

IP2

TM

EM

52

BT

ME

M2

20

TM

EM

20

6T

ME

M1

92

TM

EM

18

4B

TM

EM

14

BT

ME

D5

TM

CO

3T

LR

4T

INF

2T

ICA

M1

TH

TP

AT

HE

MIS

2T

GS

1T

GO

LN

2T

GF

BR

1T

FE

3T

CP

1T

BX

AS

1T

AR

BP

2T

AF

9B

TA

F1

AT

AD

A2

AS

YN

RG

SY

KS

WT

1S

WA

P7

0S

US

D1

SU

PT

5H

SU

GP

1S

TY

XL

1S

TK

40

ST

K1

7B

ST

AT

5A

ST

AR

D5

ST

AR

D3

NL

ST

AB

1S

T8

SIA

4S

T3

GA

L6

SS

R1

SR

SF

5S

RG

NS

RG

AP

2B

SP

P1

SP

IDR

SP

14

0S

P1

10

SP

10

0S

NX

17

SN

X1

0S

NO

RD

58

CS

NO

RA

75

SN

OR

A5

5S

NO

RA

16

AS

NO

RA

10

0S

NH

G1

5S

NC

AS

MP

D1

SL

CO

2B

1S

LC

7A

8S

LC

7A

7S

LC

7A

5S

LC

48

A1

SL

C4

7A

1S

LC

40

A1

SL

C3

9A

4S

LC

37

A2

SL

C3

5F

6S

LC

31

A2

SL

C3

1A

1S

LC

30

A4

SL

C2

9A

1S

LC

25

A2

4S

LC

1A

3S

LC

16

A1

SL

C1

5A

3S

LA

MF

8S

LA

SK

AP

2S

IRP

AS

IGL

EC

9S

IGL

EC

14

SH

Q1

SH

3D

19

SH

2B

3S

GK

1S

FT

2D

1S

ET

D3

SE

RP

INB

9S

ER

PIN

B1

SE

RP

INA

1S

EM

A4

DS

EL

PL

GS

EL

1L

3S

EL

1L

SD

SS

DC

BP

SD

C3

SC

RN

1S

CO

2S

CN

N1

BS

CD

SA

TB

1S

AT

1S

AR

1B

SA

P1

8S

AM

D9

LS

1P

R2

S1

00

A9

S1

00

A5

RT

FD

C1

RR

M2

BR

PS

19

BP

1R

PP

14

RN

VU

1-2

0R

NF

21

6P

1R

NF

14

9R

NF

14

4B

RN

AS

ET

2R

NA

SE

6R

NA

SE

1R

IPK

2R

HO

QR

HO

GR

HB

DF

2R

GS

2R

GS

10

RG

S1

RE

LR

EE

P5

RE

CQ

LR

CS

D1

RB

MX

2R

BM

47

RA

SS

F5

RA

SS

F4

RA

SS

F3

RA

SS

F2

RA

SG

EF

1B

RA

SA

2R

AP

2B

RA

D2

3A

RA

C2

RA

B2

6R

AB

20

RA

B1

0P

YC

AR

D-A

S1

PY

CA

RD

PX

DC

1P

TP

RE

PT

PR

CP

TP

N2

3P

TG

S2

PT

GS

1P

TG

ER

4P

SM

E2

PS

AP

PR

PS

1P

RE

X1

PR

EP

PR

DM

1P

PP

2R

5E

PP

M1

GP

PC

SP

OL

D4

PL

TP

PL

SC

R1

PL

P2

PL

IN3

PL

EK

HJ

1P

LE

KH

B2

PL

EK

PL

CG

2P

LB

D1

PL

AU

RP

LA

2G

7P

LA

2G

16

PIP

4K

2A

PIL

RA

PIK

3C

GP

IK3

AP

1P

IGF

PH

LD

A1

PE

CA

M1

PE

A1

5P

DP

1P

DG

FB

PD

E4

BP

BX

3P

AL

D1

PA

2G

4P

4P

2R

Y1

3P

2R

Y1

2O

TU

D1

OL

R1

OL

FM

L3

OG

FR

NX

T2

NR

IP1

NR

4A

2N

QO

2N

PC

2N

ME

6N

MB

NL

RP

3N

INJ

2N

GD

NN

FK

BIA

NF

E2

L2

NF

AT

C2

NE

K6

ND

UF

B2

-AS

1N

DS

T2

NC

OA

4N

CF

4N

CF

2N

CB

P1

NA

MP

TM

YO

9B

MY

CT

1M

X2

MX

1M

TP

NM

TM

R1

MS

R1

MS

4A

7M

S4

A6

AM

S4

A4

AM

RP

S1

8A

MR

PL

49

MR

PL

28

MR

PL

18

MR

C1

MR

1M

PP

1M

PE

G1

MN

DA

MM

DM

LK

LM

KL

1M

IR4

64

7M

IR1

55

HG

MIE

F2

MG

ST

2M

GC

16

27

5M

GA

T5

MF

SD

5M

FS

D2

AM

FS

D1

MF

HA

S1

ME

SD

C1

ME

D2

9M

ED

1M

CM

3A

PM

CL

1M

AP

KA

PK

3M

AP

3K

8M

AP

3K

2M

AP

1L

C3

B2

MA

P1

LC

3B

MA

NB

AM

AN

2B

1M

AL

T1

MA

FF

MA

FB

MA

FL

YZ

LY

NL

Y9

6L

Y8

6L

SM

5L

RR

C4

0L

RR

C2

5L

PA

R5

LP

AR

2L

OC

14

54

74

LO

C1

05

37

14

14

LO

C1

05

37

05

26

LO

C1

02

72

42

97

LO

C1

00

99

68

42

LO

C1

00

50

66

06

LO

C1

00

28

87

98

LO

C1

00

13

03

57

LO

C1

00

12

90

34

LM

O2

LIT

AF

LIP

AL

INC

01

27

2L

INC

01

12

5L

INC

01

09

4L

ILR

B5

LIL

RB

2L

GM

NL

GA

LS

9L

CP

2L

CP

1L

BR

LA

T2

LA

PT

M5

LA

CT

BL

3M

BT

L4

-AS

1K

LF

4K

LF

2K

IF3

BK

DM

2A

KC

TD

12

KA

NT

RJ

UN

BJ

MJ

D6

JM

JD

1C

JA

ZF

1J

AM

LIT

PK

1IT

GB

3IT

GB

2IT

GA

XIT

GA

MIS

CU

IRF

5IR

AK

4IQ

GA

P2

INS

IG1

INIP

IL6

IL1

BIL

18

IKZ

F1

IGS

F6

IGS

F2

1IF

IH1

IFI4

4L

IFI3

0IF

I27

L2

IFI2

7IE

R5

ID2

ICA

M1

HS

PA

7H

SP

90

AA

1H

PG

DS

HN

RN

PD

LH

NR

NP

CH

MG

N4

HM

GB

2H

MG

A1

HM

13

HL

A-D

RB

5H

LA

-DQ

A1

HK

3H

IPK

3H

IF1

A-A

S2

HE

XB

HE

XA

HE

S6

HE

RP

UD

1H

CS

TH

CL

S1

HC

KH

CA

R3

HC

AR

2H

BE

GF

HA

VC

R2

GY

PC

GS

TO

1G

SK

IPG

RW

D1

GR

NG

RA

SP

GP

X1

GP

R6

5G

PR

34

GP

R1

83

GP

R1

37

BG

PR

13

2G

PN

MB

GP

N2

GP

KO

WG

OL

GA

3G

NP

DA

1G

NG

5G

NB

1G

NA

I2G

NA

13

GM

FG

GL

UL

GL

DN

GK

GIM

AP

4G

GT

A1

PG

DA

P2

GB

P3

GB

P2

GA

TS

GA

S7

GA

PT

GA

PL

INC

GA

LT

FY

BF

US

FU

OM

FT

H1

P3

FT

H1

FP

R3

FP

R1

FO

LR

2F

NIP

2F

LII

FG

L2

FG

D2

FE

Z2

FE

RM

T3

FC

HO

2F

CG

R3

BF

CG

R3

AF

CG

R2

CF

CG

R2

BF

CG

R2

AF

CG

R1

CP

FC

GR

1B

FC

GR

1A

FC

ER

1G

FA

M4

9A

FA

M1

88

AF

AM

17

5B

FA

M1

29

AF

AM

12

0A

FA

M1

02

BE

VI2

BE

TS

2E

PS

15

EP

B4

1L

3E

NT

PD

1E

NG

EM

P3

EM

BE

LL

2E

IF2

B4

EG

R2

EB

AG

9D

YN

LR

B1

DU

SP

2D

US

P1

0D

US

P1

DP

EP

2D

OK

3D

OC

K8

DO

CK

7D

OC

K2

DN

TT

IP2

DN

AJ

C5

DIA

PH

2-A

S1

DH

OD

HD

ER

AD

EN

ND

2D

DE

F6

DA

ZA

P2

DA

PP

1D

AB

2C

YT

H1

CY

P2

S1

CY

LD

CY

FIP

1C

YB

BC

YB

AC

YB

5R

4C

XC

R4

CX

CL

8C

XC

L3

CX

CL

2C

XC

L1

6C

TS

SC

TS

DC

TS

BC

TN

SC

ST

3C

SR

NP

1C

SF

3R

CS

F2

RB

CS

F1

RC

RY

L1

CR

Y1

CR

EM

CR

EB

L2

CP

VL

CP

SF

2C

PE

D1

CO

X5

AC

OX

14

CO

TL

1C

OM

MD

9C

NP

Y3

CM

TM

7C

MK

LR

1C

LE

C7

AC

LE

C4

AC

LD

N1

CK

LF

CID

EB

CH

CH

D1

0C

EP

63

CE

P3

50

CE

P1

70

CE

CR

1C

DC

73

CD

93

CD

86

CD

84

CD

83

CD

81

CD

74

CD

69

CD

68

CD

53

CD

40

CD

4C

D3

7C

D3

3C

D2

8C

D1

63

CD

14

CC

R1

CC

L8

CC

L4

L2

CC

L4

L1

CC

L4

CC

L3

L3

CC

L3

L1

CC

L3

CC

L2

0C

CL

2C

CD

C8

8A

CC

DC

11

2C

AS

P1

0C

AS

P1

CA

RD

8C

AR

D1

6C

AP

GC

AB

39

C9

orf7

2C

6o

rf62

C5

AR

1C

3A

R1

C3

C2

orf6

9C

1Q

CC

1Q

BC

1Q

AC

1o

rf16

2C

17

orf5

8C

15

orf4

8B

OR

CS

8B

MP

2K

BL

VR

BB

LO

C1

S2

BL

NK

BIR

C3

BID

BC

L2

L1

BC

AT

1B

CA

S2

BA

Z1

AA

TP

8B

4A

TP

6V

1F

AT

P6

V1

B2

AT

P6

V0

BA

TP

13

A3

AT

F3

AS

NS

D1

AS

LA

RR

B2

AR

PC

5L

AR

PC

3A

RP

C1

BA

RL

6IP

4A

RL

4C

AR

ID4

AA

RH

GA

P1

8A

RH

GA

P1

5A

PR

TA

PO

C2

AP

OC

1A

PH

1B

AP

CA

PB

B1

IPA

P2

S1

AP

1B

1A

NK

RD

44

AN

KF

Y1

AN

GA

MZ

2A

MD

HD

2A

LO

X5

AP

AL

OX

5A

LD

H3

B2

AL

CA

MA

KR

1B

1A

KA

P1

3A

DP

RH

AD

GR

E2

AD

AP

2A

DA

M2

8A

CT

R3

AC

SL

1A

CR

BP

AC

AA

2A

BT

1A

BR

AC

LA

BC

F2

ZN

F8

29

ZN

F8

ZN

F7

73

ZN

F7

21

ZN

F6

84

ZN

F6

52

ZN

F6

39

ZN

F6

05

ZN

F4

84

ZN

F4

68

ZN

F4

51

ZN

F4

38

ZN

F4

19

ZN

F3

94

ZN

F3

33

ZN

F3

24

ZN

F3

0Z

NF

28

0D

ZN

F2

77

ZN

F1

38

ZH

X1

ZF

YV

E1

9Z

FP

90

ZF

HX

4Z

EB

2Z

EB

1Z

C4

H2

ZC

3H

7A

ZB

TB

39

ZB

TB

38

YW

HA

EY

IF1

AY

AE

1D

1X

RR

A1

XA

F1

WW

C3

WT

AP

WN

T5

AW

ISP

1W

HA

MM

WD

R4

5W

AS

HC

2A

VS

TM

4V

RK

3V

PS

8V

PS

13

BV

OP

P1

VIM

VH

LV

GL

L3

VE

GF

BV

DA

C3

VA

MP

5U

SP

9X

US

P7

US

P3

1U

SP

3U

SP

22

US

P2

US

P1

3U

RO

DU

QC

R1

1U

NC

5B

UG

GT

1U

FL

1U

BX

N4

UB

L5

UB

E4

AU

BE

2Q

2P

1U

BE

2F

UB

E2

E2

UB

E2

D4

UB

E2

D3

TX

NR

D1

TW

SG

1T

WIS

T1

TU

BB

6T

TL

L3

TT

C3

7T

ST

A3

TS

R2

TS

PA

N6

TS

PA

N3

1T

SG

10

1T

SE

N1

5T

SC

22

D3

TS

C2

2D

1T

RIM

8T

RIM

59

TR

IM2

2T

RIL

TR

AP

PC

5T

RA

PP

C2

BT

RA

2A

TP

T1

TP

PP

3T

PM

1T

PB

GT

P5

3IN

P1

TO

R1

AIP

1T

OM

M7

TN

S1

TN

RC

6B

TN

KS

TN

FS

F1

0T

NF

RS

F2

1T

NF

RS

F1

2A

TN

FA

IP6

TM

X1

TM

LH

ET

MF

1T

ME

M5

9T

ME

M4

5A

TM

EM

35

BT

ME

M3

0A

TM

EM

25

1T

ME

M2

48

TM

EM

21

9T

ME

M2

12

TM

EM

20

4T

ME

M2

00

AT

ME

M1

4A

TM

EM

12

6B

TM

EM

12

0A

TM

EM

11

TM

ED

4T

MB

IM6

TM

BIM

4T

LE

1T

JP

1T

IMP

3T

IMP

2T

IMP

1T

IMM

9T

IMM

22

TH

Y1

TH

UM

PD

1T

HR

AP

3T

HO

C2

TH

BS

4T

HB

S2

TH

AP

4T

GF

BR

2T

GF

BI

TG

FB

3T

FP

IT

FB

1M

TF

AM

TE

X2

61

TD

P2

TC

F4

TC

EA

L9

TC

EA

L4

TB

X1

5T

BR

G1

TB

CB

TB

C1

D2

4T

BC

1D

15

TA

GL

NT

AF

8T

AF

5L

TA

DA

2B

TA

CC

1S

YT

11

SY

NP

O2

SY

NJ

2B

P-C

OX

16

SY

NJ

2B

PS

YN

GR

1S

YN

E2

SY

NE

1-A

S1

SW

I5S

UM

O3

SU

MO

2S

UL

F1

SU

GT

1S

UC

LA

2S

TX

7S

TX

2S

TT

3A

ST

RN

3S

TR

AP

ST

N1

ST

K3

ST

EA

P2

ST

EA

P1

ST

AT

1S

T7

LS

ST

R2

SS

SC

A1

SS

R4

SS

PN

SS

BS

RP

X2

SR

PX

SR

P9

SR

P5

4S

RG

AP

2D

SR

EK

1IP

1S

QS

TM

1S

QR

DL

SP

TB

N1

SP

RY

4-IT

1S

PO

PS

PO

N2

SP

OC

K1

SP

IN1

SP

HK

1S

PG

20

SP

CS

3S

PC

S2

SP

C2

5S

PA

TS

2L

SP

AR

CL

1S

PA

RC

SN

X6

SN

X2

SN

U1

3S

NR

PN

SN

RN

P3

5S

NO

RA

10

4S

NE

D1

SN

AP

C1

SN

AP

23

SM

OC

2S

MIM

10

L1

SM

C6

SM

C1

AS

MA

RC

A1

SM

AP

1S

LIT

2S

LC

7A

6S

LC

6A

6S

LC

5A

3S

LC

50

A1

SL

C4

4A

1S

LC

41

A2

SL

C3

5F

5S

LC

35

F1

SL

C3

5B

4S

LC

30

A7

SL

C2

A3

SL

C2

A1

3S

LC

26

A2

SL

C1

2A

8S

IPA

1L

1S

H3

KB

P1

SH

3G

LB

1S

GP

P1

SG

MS

2S

GIP

1S

GF

29

SG

CE

SG

CB

SF

RP

4S

FR

P2

SE

RP

ING

1S

ER

PIN

E1

SE

RP

INB

6S

ER

P1

SE

RF

2S

EN

P6

SE

MA

5A

SE

LE

NO

WS

EL

EN

ON

SE

C2

4D

SE

C2

3A

SE

C2

2B

SD

HB

SD

F2

L1

SD

F2

SD

C2

SC

N7

AS

CM

L1

SC

AR

NA

18

SB

NO

1S

BD

SP

1S

BD

SS

AR

SS

AR

NP

SA

P3

0B

PS

AM

HD

1S

AF

BS

10

0A

6S

10

0A

3R

WD

D3

RU

NX

1T

1R

RP

1B

RR

P1

5R

RB

P1

RR

AG

BR

PS

15

AP

10

RP

P3

0R

PE

RO

R2

RN

PC

3R

NF

7R

NF

21

6R

NF

18

5R

NF

17

0R

NF

13

5R

NF

13

0R

NF

11

1R

NF

11

RN

AS

EL

RN

AS

EK

RIT

A1

RIN

2R

HO

BT

B3

RH

BD

D3

RG

S5

RG

S1

6R

GC

CR

FT

N2

RF

LN

BR

EL

L1

RE

EP

3R

DX

RD

H1

1R

BP

JR

BM

S3

RB

M6

RB

M3

RB

M1

7R

BA

KR

B1

CC

1R

AS

SF

8R

AR

RE

S2

RA

RB

RA

PG

EF

2R

AP

1G

DS

1R

AI2

RA

I14

RA

D5

1B

RA

B5

BR

AB

40

BR

AB

3G

AP

2R

AB

22

AR

AB

21

RA

B1

5R

AB

12

PW

P1

PU

RA

PU

M3

PT

RF

PT

PN

12

PT

P4

A3

PT

NP

TK

2P

TG

R1

PT

GIS

PT

GF

RN

PT

EN

PT

BP

2P

SM

E1

PS

MD

2P

SM

C2

PS

MB

4P

SM

A3

PS

MA

1P

SE

NE

NP

RS

S2

3P

RR

X1

PR

RG

1P

RO

S1

PR

MT

7P

RK

D1

PR

KC

DB

PP

RA

F2

PQ

LC

3P

PP

6C

PP

P4

R3

BP

PP

1R

12

AP

PE

F1

PO

ST

NP

ON

2P

OM

T1

PO

LR

2K

PO

LL

PO

GL

UT

1P

MS

2P

3P

MP

22

PM

EP

A1

PL

XN

D1

PL

XN

C1

PL

XD

C1

PL

SC

R4

PL

S3

PL

PP

1P

LN

PL

IN2

PL

EK

HO

2P

LC

E1

PL

AU

PL

AG

L1

PL

AC

9P

LA

2G

5P

KN

2P

KM

PK

IGP

KD

2P

JA

2P

IP4

K2

CP

INK

1-A

SP

INK

1P

IGV

PIG

CP

HT

F1

PH

LD

B1

PH

AX

PG

M3

PG

BD

1P

FD

N4

PE

LO

PE

LI2

PE

AR

1P

DZ

D1

1P

DS

5B

PD

LIM

3P

DIA

3P

DG

FR

BP

DG

FR

AP

DE

5A

PD

E3

AP

DE

1A

PD

CD

5P

CO

LC

EP

AT

L1

PA

RP

BP

PA

RP

9P

AR

P8

PA

RP

4P

AR

P1

4P

4H

A1

OX

CT

1O

TU

D6

B-A

S1

OS

T4

OS

R2

OS

MR

OS

BP

L9

OR

C4

OR

51

E1

OR

2A

7O

PH

N1

OM

DO

MA

1O

LF

ML

1O

LF

M1

OG

TO

DF

2O

CE

L1

OA

FN

UT

M2

A-A

S1

NU

TF

2N

UP

L2

NU

DT

10

NU

CB

2N

UB

1N

TM

NT

5C

NR

P1

NR

EP

NR

1D

2N

PT

N-IT

1N

OT

CH

3N

OR

AD

NO

C3

LN

NM

TN

MI

NL

GN

4Y

NK

AP

NIP

AL

2N

ID2

NID

1N

FK

BIZ

NF

2N

EIL

2N

DU

FV

3N

DU

FC

2N

DU

FB

10

ND

UF

AF

1N

DU

FA

6N

DU

FA

4L

2N

DU

FA

3N

DU

FA

13

ND

UF

A1

2N

DU

FA

1N

CO

R1

NC

OA

1N

BR

1N

BP

F8

NB

PF

15

NB

NN

BL

1N

BE

AL

1N

AV

1N

AP

1L

3N

AG

KN

AD

KN

AA

20

NA

A1

0N

4B

P2

L2

MY

O1

BM

YO

19

MY

NN

MY

H9

MX

RA

8M

XR

A5

MT

ER

F4

MT

CH

1M

RV

I1M

RP

S1

8C

MR

PS

15

MR

PS

11

MR

PL

51

MR

PL

33

MR

PL

2M

RP

L1

5M

RP

L1

4M

PR

IPM

PP

5M

PD

U1

MP

C1

MO

RN

2M

OR

F4

L1

MO

RC

4M

NS

1M

MP

2M

MP

16

MM

P1

1M

LE

CM

KL

N1

-AS

MIR

99

AH

GM

IR5

04

7M

IR4

43

5-2

HG

MIR

36

52

MIN

CR

MIE

R1

MG

ST

3M

FN

2M

FA

P4

ME

TT

L4

ME

TT

L2

3M

ET

TL

17

ME

TA

P2

ME

ST

ME

SD

C2

ME

F2

CM

ED

4-A

S1

ME

D4

ME

D1

2M

DP

1M

DF

ICM

CU

BM

CM

6M

CF

D2

MC

AM

MB

NL

1M

BD

3M

AP

RE

2M

AP

K1

IP1

LM

AP

4K

4M

AP

4M

AP

1B

MA

P1

AM

AN

1A

1M

AL

SU

1M

AG

T1

MA

GO

HB

MA

GO

HM

AG

I3M

AF

G-A

S1

MA

D2

L1

BP

LY

RM

2L

YP

LA

L1

LY

AR

LX

NL

UM

LT

BP

2L

SM

3L

SG

1L

RR

C5

7L

RR

C3

2L

RR

C1

5L

RP

10

LR

P1

LR

IG2

LR

CH

3L

OX

LO

C4

00

04

3L

OC

28

40

23

LO

C1

43

66

6L

OC

10

53

71

81

4L

OC

10

27

24

55

2L

OC

10

19

29

34

0L

OC

10

19

28

13

7L

OC

10

19

28

09

3L

OC

10

19

27

31

1L

OC

10

05

06

38

8L

OC

10

04

19

17

0L

OC

10

01

31

28

9L

INC

01

42

0L

INC

00

98

9L

INC

00

96

3L

IN3

7L

IMS

1L

HF

PL

GA

LS

LL

EP

RO

TL

1L

EP

RO

TL

EP

RL

EN

G1

LB

HL

AM

TO

R3

LA

MT

OR

1L

AM

P5

LA

MP

2L

AM

A4

L3

MB

TL

3K

RB

OX

4K

MT

5A

KM

T2

CK

LF

10

KID

INS

22

0K

IAA

15

51

KIA

A0

04

0K

DE

LR

3K

CT

D2

0K

CT

D1

5K

CN

S3

KA

TN

BL

1K

AT

6A

KA

RS

KA

NK

2J

AK

1J

AG

1IT

PR

IPL

2IT

M2

CIT

M2

BIT

GB

L1

ITG

B5

ITG

B1

BP

1IT

GB

1IT

GA

VIT

GA

EIT

GA

1IT

FG

2IS

LR

IRF

7IR

F2

BP

2IN

TU

INT

S1

0IN

PP

4B

INH

BA

IL3

2IL

1R

1IG

FB

P7

IGF

BP

5IG

FB

P4

IGF

BP

3IG

F1

IGD

CC

4IF

T5

2IF

T2

7IF

NG

R1

IFIT

M3

IFIT

M1

IFI1

6ID

SID

H3

AID

3H

YP

KH

UW

E1

HT

RA

3H

SP

H1

HS

PB

AP

1H

SP

B1

HS

D1

7B

7H

SD

17

B1

1H

SD

17

B1

0H

PR

T1

HP

F1

HO

TA

IRH

OP

XH

OO

K3

HN

RN

PU

HN

RN

PR

HN

RN

PH

2H

NR

NP

A0

HM

CN

1H

LT

F-A

S1

HIV

EP

3H

IPK

1H

INT

2H

IGD

1B

HG

SN

AT

HE

YL

HE

G1

HD

GF

RP

3H

DD

C2

HD

AC

9H

DA

C8

HC

FC

2H

CF

C1

R1

HA

RS

HA

AO

GX

YL

T1

GU

SB

GU

CY

1B

3G

UC

Y1

A3

GU

CD

1G

TF

2H

5G

TF

2E

1G

TF

2B

GS

TM

3G

ST

A4

GS

NG

RP

GP

X8

GP

X4

GP

R6

8G

PR

17

6G

PC

1G

OL

IM4

GN

G1

2G

NG

11

GN

A1

2G

LU

D1

GL

T8

D2

GL

IPR

2G

LG

1G

JC

1G

INM

1G

GT

5G

GN

BP

2G

BP

1G

AL

3S

T4

G3

BP

2F

XY

D6

FU

T8

FT

OF

ST

L1

FR

ZB

FR

Y-A

S1

FR

YF

RM

D6

FR

KF

N1

FM

OD

FM

O3

FL

RT

2F

LJ

20

02

1F

KB

P9

P1

FK

BP

9F

KB

P7

FK

BP

11

FIS

1F

ILIP

1L

FIB

INF

HL

1F

GF

R1

OP

2F

GD

5-A

S1

FC

GR

TF

BX

O7

FB

XO

42

FB

XL

5F

BN

1F

BL

N5

FB

LN

1F

AT

4F

AS

FA

PF

AN

CG

FA

M8

A1

FA

M2

29

BF

AM

20

CF

AM

20

8A

FA

M2

00

BF

AM

19

9X

FA

M1

98

BF

AM

17

9B

FA

M1

68

AF

AM

13

CF

AM

11

4A

1F

AM

11

1A

FA

M1

10

BF

AH

FA

BP

5E

XT

1E

XO

SC

9E

XO

SC

8E

TV

1E

TN

K1

ES

AM

ER

VK

3-1

ER

O1

BE

RL

IN2

ER

HE

RG

IC2

ER

AP

2E

PS

8E

PR

SE

PB

41

L2

EP

AS

1E

MP

2E

ML

4E

MC

10

EL

OV

L5

EL

OB

EL

K4

EL

F1

EIF

5B

EIF

5A

2E

IF4

G3

EIF

4E

EIF

3I

EIF

3E

EIF

2S

3E

IF2

S1

EIF

2B

3E

HD

2E

GL

N3

EF

TU

D2

EF

EM

P2

EE

PD

1E

EA

1E

DN

RA

ED

EM

1E

CM

2E

BL

N3

PE

BF

1D

YN

LL

1D

YN

C1

LI1

DT

X4

DS

TD

SE

LD

PY

SL

3D

PY

SL

2D

PY

DD

OP

EY

1D

OC

K5

DO

CK

1D

NA

SE

1L

1D

NA

JC

15

DN

AJ

C1

3D

NA

JB

4D

NA

JB

11

DL

GA

P1

-AS

1D

LD

DL

C1

DIS

3D

IO2

DID

O1

DIA

PH

2D

HR

S4

DE

GS

1D

EF

8D

DX

3Y

DD

X3

9A

DD

R2

DD

B2

DC

ND

CB

LD

2D

CA

F5

DB

ND

D2

DA

D1

CY

TO

RC

YR

61

CY

P5

1A

1C

YB

RD

1C

XC

L1

2C

TS

KC

TP

S2

CT

HR

C1

CT

DS

P1

CT

CF

CS

TF

1C

SP

P1

CR

Y2

CR

TA

PC

RIS

PL

D2

CR

IPT

CR

IP2

CR

IP1

CP

EC

OX

4I1

CO

X1

7C

OP

Z2

CO

PS

8C

OP

S5

CO

MT

CO

L8

A1

CO

L6

A3

CO

L6

A2

CO

L6

A1

CO

L5

A2

CO

L5

A1

CO

L4

A1

CO

L3

A1

CO

L1

A2

CO

L1

A1

CO

L1

8A

1C

OL

14

A1

CO

L1

2A

1C

OL

11

A1

CO

L1

0A

1C

OG

3C

OA

1C

NR

IP1

CN

PC

NN

3C

NIH

4C

NE

P1

R1

CM

TR

2C

MT

M3

CM

AH

PC

LE

C2

BC

LC

N5

CL

CN

3C

IR1

CH

UR

C1

CH

ST

3C

HO

RD

C1

CH

CH

D2

CF

HR

1C

FH

CE

P1

12

CE

NP

CC

DK

L1

CD

K5

CD

K3

CD

K1

4C

DH

6C

DH

13

CD

H1

1C

DC

5L

CD

C4

2B

PA

CD

C3

7L

1C

DC

26

CD

99

CD

63

CD

30

2C

D2

48

CD

16

0C

D1

09

CC

Z1

BC

CZ

1C

CN

IC

CM

2C

CD

C8

2C

CD

C8

0C

CD

C5

1C

CD

C2

5C

CD

C1

74

CB

X6

CB

WD

6C

BW

D3

CA

V2

CA

V1

CA

SC

4C

AP

ZB

CA

PN

2C

AP

1C

AM

K2

N1

CA

MK

1D

CA

LU

CA

LR

CA

LD

1C

AD

M1

C8

orf4

4C

7o

rf26

C6

orf8

9C

6o

rf10

6C

4o

rf3C

3o

rf80

C3

orf7

0C

3o

rf38

C3

orf1

4C

20

orf2

7C

1S

C1

RC

1Q

TN

F5

C1

5o

rf39

C1

2o

rf10

C1

0o

rf10

BT

N3

A2

BT

N2

A1

BR

OX

BR

K1

BM

P8

AB

LO

C1

S6

BIC

C1

BG

NB

EX

3B

ET

1L

BC

AP

29

BA

NF

1B

4G

AL

T3

B4

GA

LT

1A

XL

AT

RA

IDA

TP

6V

1C

1A

TP

5J

AT

P5

F1

AT

P5

C1

AT

OX

1A

TL

3A

TL

1A

TG

10

AT

F6

AT

F2

AS

PN

AS

H2

LA

RP

C4

AR

MC

X3

AR

L2

AR

L1

AR

ID5

BA

RH

GA

P5

AR

HG

AP

32

AR

HG

AP

29

AR

HG

AP

28

AR

HG

AP

17

AR

HG

AP

1A

RF

GE

F1

AR

F6

AR

F5

AP

PL

2A

PO

L2

AP

OL

1A

PO

DA

PC

DD

1A

P3

B1

AN

XA

7A

NX

A5

AN

XA

4A

NX

A1

1A

NT

XR

2A

NT

XR

1A

NO

1A

NK

RD

10

AN

KH

AN

GP

TL

1A

NG

PT

2A

NA

PC

10

AM

PD

2A

LK

BH

5A

LG

13

AL

DH

1L

2A

LD

H1

A3

AL

DH

1A

1A

JU

BA

AID

AA

HN

AK

AG

PA

T5

AE

BP

1A

DIR

FA

DG

RF

5A

DC

Y3

AD

AM

TS

L3

AD

AM

TS

4A

DA

M9

AD

AM

19

AD

AA

CV

R1

BA

CT

N1

AC

LY

AC

AD

11

AB

HD

18

AB

HD

10

AB

CC

9A

BC

A9

AB

CA

6A

2M

A1

BG

43

35

44

33

53

43

35

04

33

47

43

16

5Y

AP

1W

DR

6W

BP

1V

ITV

CA

NU

SP

34

TU

FM

TT

C1

9T

ST

D1

TR

IM2

TM

4S

F1

TG

FB

R3

TF

AP

2C

TC

F7

L1

TC

ER

G1

TB

X3

SR

PK

2S

OX

11

SO

RB

S2

SN

X2

2S

NR

NP

40

SN

HG

3S

NH

G2

5S

MIM

4S

MA

D5

SM

A4

SIK

1S

HC

4S

FR

P1

SE

TD

5S

ER

PIN

A3

SC

UB

E3

SC

RG

1S

CN

N1

GS

CH

IP1

SA

A2

SA

A1

RT

F1

RS

F1

RP

S2

1R

PL

7A

RP

L1

1R

PA

INR

NP

S1

RB

PM

SR

AR

SP

YG

BP

UM

1P

TP

RF

PR

KY

PR

KC

IP

RE

LP

PP

M1

KP

NIS

RP

IK3

R3

PIK

3R

1P

HK

G1

PG

AP

1P

ER

2P

EG

3-A

S1

PE

G3

PD

E9

AP

CM

TD

1P

AR

D6

BP

AR

D3

PA

PS

S1

PA

DI2

OB

P2

BO

BP

2A

NU

DT

5N

RP

2N

PIP

B5

NP

IPB

4N

PIP

B3

NP

IPB

13

NP

IPB

11

NIP

AL

3N

FIB

ND

UF

A7

NA

V2

MZ

T2

AM

YC

MT

HF

D2

MT

2A

MP

ZL

1M

LX

IPM

IAM

GS

T1

ME

IS1

MB

NL

2M

AR

CK

SL

1M

AL

AT

1M

AE

AM

AB

21

L3

LIN

C0

12

96

KR

T8

KR

T5

KR

T2

3K

RT

19

KR

T1

8K

RT

15

KR

T1

4K

LF

5K

DS

RK

AN

K4

JU

PIT

GB

4IT

GA

6IS

G2

0L

2IR

F6

IGF

2IG

F1

RH

P1

BP

3H

19

GT

F2

IP1

GR

HL

1G

PA

TC

H8

GO

LP

H3

GA

S5

GA

BR

PF

ZD

7F

XY

D3

FR

MD

4A

FL

NB

FG

FR

2F

GF

R1

FB

XO

32

FB

LF

AT

1F

AM

60

AF

AB

P7

ES

RP

1E

RR

FI1

EP

CA

ME

HF

EG

FR

EF

NA

1D

SP

DS

G2

DS

C3

DS

C2

CX

AD

RC

TB

P2

CS

TF

3C

RIM

1C

RA

BP

2C

OL

9A

2C

OL

9A

1C

OL

27

A1

CM

YA

5C

LD

N4

CD

K6

CD

H1

CD

24

CC

NT

2C

CN

L2

CA

PN

12

C2

1o

rf91

BO

CB

CL

11

AB

AM

BI

AT

P5

A1

AT

F4

AR

T3

AR

ID1

AA

QP

1A

PP

AN

XA

3A

NK

RD

36

AK

AP

9A

DN

PA

CT

G2

04

81

2

Va

lue

0 60000

Co

lor K

ey

an

d H

isto

gra

m

Count


Tum

or_

C91

Tum

or_

C75

Tum

or_

C34

Tum

or_

C19

Tum

or_

C12

Tum

or_

C10

Tum

or_

C69

Tum

or_

C53

Tum

or_

C42

Tum

or_

C41

Tum

or_

C26

Tum

or_

C04

Tum

or_

C01

Tum

or_

C95

Tum

or_

C94

Tum

or_

C93

Tum

or_

C92

Tum

or_

C90

Tum

or_

C89

Tum

or_

C88

Tum

or_

C87

Tum

or_

C86

Tum

or_

C85

Tum

or_

C84

Tum

or_

C82

Tum

or_

C81

Tum

or_

C79

Tum

or_

C77

Tum

or_

C76

Tum

or_

C74

Tum

or_

C73

Tum

or_

C72

Tum

or_

C71

Tum

or_

C70

Tum

or_

C68

Tum

or_

C66

Tum

or_

C63

Tum

or_

C62

Tum

or_

C60

Tum

or_

C58

Tum

or_

C57

Tum

or_

C51

Tum

or_

C50

Tum

or_

C49

Tum

or_

C43

Tum

or_

C40

Tum

or_

C38

Tum

or_

C37

Tum

or_

C33

Tum

or_

C32

Tum

or_

C28

Tum

or_

C27

Tum

or_

C24

Tum

or_

C23

Tum

or_

C22

Tum

or_

C20

Tum

or_

C16

Tum

or_

C15

Tum

or_

C14

Tum

or_

C08

Tum

or_

C07

Tum

or_

C06

Tum

or_

C05

Tum

or_

C02

0.2

0.6

1

Va

lue

0 200

Co

lor K

ey

an

d H

isto

gra

m

Count

Tum

or_

C02

Tum

or_

C05

Tum

or_

C06

Tum

or_

C07

Tum

or_

C08

Tum

or_

C14

Tum

or_

C15

Tum

or_

C16

Tum

or_

C20

Tum

or_

C22

Tum

or_

C23

Tum

or_

C24

Tum

or_

C27

Tum

or_

C28

Tum

or_

C32

Tum

or_

C33

Tum

or_

C37

Tum

or_

C38

Tum

or_

C40

Tum

or_

C43

Tum

or_

C49

Tum

or_

C50

Tum

or_

C51

Tum

or_

C57

Tum

or_

C58

Tum

or_

C60

Tum

or_

C62

Tum

or_

C63

Tum

or_

C66

Tum

or_

C68

Tum

or_

C70

Tum

or_

C71

Tum

or_

C72

Tum

or_

C73

Tum

or_

C74

Tum

or_

C76

Tum

or_

C77

Tum

or_

C79

Tum

or_

C81

Tum

or_

C82

Tum

or_

C84

Tum

or_

C85

Tum

or_

C86

Tum

or_

C87

Tum

or_

C88

Tum

or_

C89

Tum

or_

C90

Tum

or_

C92

Tum

or_

C93

Tum

or_

C94

Tum

or_

C95

Tum

or_

C01

Tum

or_

C04

Tum

or_

C26

Tum

or_

C41

Tum

or_

C42

Tum

or_

C53

Tum

or_

C69

Tum

or_

C10

Tum

or_

C12

Tum

or_

C19

Tum

or_

C34

Tum

or_

C75

Tum

or_

C91


0.2 0.6 1

Value

02

00

Color Key

and Histogram

Count

Tum

or_

C02

Tum

or_

C05

Tum

or_

C06

Tum

or_

C07

Tum

or_

C08

Tum

or_

C14

Tum

or_

C15

Tum

or_

C16

Tum

or_

C20

Tum

or_

C22

Tum

or_

C23

Tum

or_

C24

Tum

or_

C27

Tum

or_

C28

Tum

or_

C32

Tum

or_

C33

Tum

or_

C37

Tum

or_

C38

Tum

or_

C40

Tum

or_

C43

Tum

or_

C49

Tum

or_

C50

Tum

or_

C51

Tum

or_

C57

Tum

or_

C58

Tum

or_

C60

Tum

or_

C62

Tum

or_

C63

Tum

or_

C66

Tum

or_

C68

Tum

or_

C70

Tum

or_

C71

Tum

or_

C72

Tum

or_

C73

Tum

or_

C74

Tum

or_

C76

Tum

or_

C77

Tum

or_

C79

Tum

or_

C81

Tum

or_

C82

Tum

or_

C84

Tum

or_

C85

Tum

or_

C86

Tum

or_

C87

Tum

or_

C88

Tum

or_

C89

Tum

or_

C90

Tum

or_

C92

Tum

or_

C93

Tum

or_

C94

Tum

or_

C95

Tum

or_

C01

Tum

or_

C04

Tum

or_

C26

Tum

or_

C41

Tum

or_

C42

Tum

or_

C53

Tum

or_

C69

Tum

or_

C10

Tum

or_

C12

Tum

or_

C19

Tum

or_

C34

Tum

or_

C75

Tum

or_

C91

Tumor_C20Tumor_C16Tumor_C15Tumor_C14Tumor_C08Tumor_C07Tumor_C06Tumor_C05Tumor_C02

figure margins too large

Cluster_1

Cluster_2

Cluster_3

Tum

or_

C02

Tum

or_

C05

Tum

or_

C06

Tum

or_

C07

Tum

or_

C08

Tum

or_

C14

Tum

or_

C15

Tum

or_

C16

Tum

or_

C20

Tum

or_

C22

Tum

or_

C23

Tum

or_

C24

Tum

or_

C27

Tum

or_

C28

Tum

or_

C32

Tum

or_

C33

Tum

or_

C37

Tum

or_

C38

Tum

or_

C40

Tum

or_

C43

Tum

or_

C49

Tum

or_

C50

Tum

or_

C51

Tum

or_

C57

Tum

or_

C58

Tum

or_

C60

Tum

or_

C62

Tum

or_

C63

Tum

or_

C66

Tum

or_

C68

Tum

or_

C70

Tum

or_

C71

Tum

or_

C72

Tum

or_

C73

Tum

or_

C74

Tum

or_

C76

Tum

or_

C77

Tum

or_

C79

Tum

or_

C81

Tum

or_

C82

Tum

or_

C84

Tum

or_

C85

Tum

or_

C86

Tum

or_

C87

Tum

or_

C88

Tum

or_

C89

Tum

or_

C90

Tum

or_

C92

Tum

or_

C93

Tum

or_

C94

Tum

or_

C95

Tum

or_

C01

Tum

or_

C04

Tum

or_

C26

Tum

or_

C41

Tum

or_

C42

Tum

or_

C53

Tum

or_

C69

Tum

or_

C10

Tum

or_

C12

Tum

or_

C19

Tum

or_

C34

Tum

or_

C75

Tum

or_

C91



Cluster_1

Cluster_2

Cluster_3

Tum

or_

C02

Tum

or_

C05

Tum

or_

C06

Tum

or_

C07

Tum

or_

C08

Tum

or_

C14

Tum

or_

C15

Tum

or_

C16

Tum

or_

C20

Tum

or_

C22

Tum

or_

C23

Tum

or_

C24

Tum

or_

C27

Tum

or_

C28

Tum

or_

C32

Tum

or_

C33

Tum

or_

C37

Tum

or_

C38

Tum

or_

C40

Tum

or_

C43

Tum

or_

C49

Tum

or_

C50

Tum

or_

C51

Tum

or_

C57

Tum

or_

C58

Tum

or_

C60

Tum

or_

C62

Tum

or_

C63

Tum

or_

C66

Tum

or_

C68

Tum

or_

C70

Tum

or_

C71

Tum

or_

C72

Tum

or_

C73

Tum

or_

C74

Tum

or_

C76

Tum

or_

C77

Tum

or_

C79

Tum

or_

C81

Tum

or_

C82

Tum

or_

C84

Tum

or_

C85

Tum

or_

C86

Tum

or_

C87

Tum

or_

C88

Tum

or_

C89

Tum

or_

C90

Tum

or_

C92

Tum

or_

C93

Tum

or_

C94

Tum

or_

C95

Tum

or_

C01

Tum

or_

C04

Tum

or_

C26

Tum

or_

C41

Tum

or_

C42

Tum

or_

C53

Tum

or_

C69

Tum

or_

C10

Tum

or_

C12

Tum

or_

C19

Tum

or_

C34

Tum

or_

C75

Tum

or_

C91

RPP14RPS19BP1THTPADNTTIP2AMDHD2ARID4AIRF5CDC73NGDNBCAS2BAZ1AMRPS18ABIDSNORA55RAD23ATMEM184BMED1CRYL1PXDC1WASHC4XRN2SNX17ZEB2-AS1PLP2SP100SLC40A1IRAK4FUSURB1-AS1WASH3PISCUSNORD58CNQO2SAP18STYXL1C6orf62TMEM14BSLC39A4SLC31A1NINJ2STK17BTSPAN14ZNF718TCP1GRWD1S100A5L3MBTL4-AS1ABT1AMZ2ASNSD1PPM1GSUPT5HSDC3SCRN1PTGER4SSR1FUOMRNF216P1SP110IER5GNB1HNRNPCTGS1TNFAIP2HSP90AA1FAM120AMANBALSM5GALTLOC100288798POLD4YIPF5TGFBR1CEP63HMGN4ARHGAP15BLVRBMIR4647ARPC3LRRC40SATB1TSPAN33SPIDRTGOLN2TXNDC11SETD3UTP14ASHQ1DYNLRB1MYCT1C17orf58CD33LITAFRASSF2CHCHD10RNF144BSCNN1BPTGS1COX5AMGC16275LOC100129034SNORA100FAM188AHM13SAR1BFEZ2MR1ACTR3UBXN2BACAA2DUSP2SMPD1UBE2D1ENTPD1SLC30A4CD69NRIP1BORCS8SLC48A1SLC7A8PIGFABRACLATP6V1B2SH3D19IL18IFIH1SKAP2RHOQARPC5LMRPL18MAP1LC3BMED29ASLRTFDC1SAT1CD81ST8SIA4ANKRD44CTSBHEXAP2RY13TAF9BRECQLSEL1LTPM3ATP13A3MTMR1PPP2R5ESWAP70MX2GAPLINCGSTO1HIPK3MRPL49ATF3MAP1LC3B2ZFP36L2MCM3APRASGEF1BSERPINB1EIF2B4GOLGA3GRNTADA2AAPRTTAF1AOGFRZNF281MGST2DUSP1HIF1A-AS2PIK3AP1C15orf48AP2S1DEF6RASA2AKAP13SCO2ATP6V0BITPK1MAFFYBX1SIRPAKDM2ARBMX2ATP8B4IFI27CPSF2DENND2DTMED5EGR2CYP2S1CREMADGRE2PEA15DAZAP2CTSDCLEC4AP2RY12MALT1COMMD9NPC2WIPF1UBAC1JMJD6PLEKHB2LYNCYLDSWT1COX14MFHAS1SUGP1HNRNPDLVSIG4NFE2L2MX1ATP6V1FC9orf72YWHAHPTPN23RBM47SNORA16ACRY1WFDC21PHMGB2GYPCUSF2ANGLPAR2NDST2XXYLT1MIEF2PPCSGBP3NCBP1HSPA7PDP1INSIG1MRPL28S1PR2TMEM206ALOX5GLULKIF3BLOC100506606DOCK7MTPNPBX3BIRC3FAM175BGPNMBRASSF3GATSAKR1B1TOX4GNG5PLSCR1HES6SRSF5PSAPCCDC112FLIILGMNZNF331HERPUD1ZYXLOC145474ZNF804ASNHG15PREPTARBP2LOC105370526MFSD5GAS7ALDH3B2SFT2D1CEP170TMCO3SEL1L3CD40MIR155HGCNPY3HCAR2EBAG9IFI44LNAMPTPLIN3MCL1CTNSID2LBRSYNRGDUSP10PRPS1SH2B3RNASE1NR4A2CYTH1RAP2BPIP4K2ACEP350ZBTB20-AS4FAM102BGNPDA1ADPRHVAMP8TNFSF13BEPS15PDGFBSLC16A1PSME2ACRBPRAB26SERPINA1ST3GAL6TMEM220LILRB5SLC47A1SUSD1LINC01125GLDNGPN2CD28CASP10GPKOWNDUFB2-AS1LOC100130357KANTRFTH1SLC7A5CLDN1NEK6JMJD1CLOC100996842REEP5INIPSDCBPGPR137BMESDC1GPX1ZDHHC2XYLT1OLFML3GRASPBLOC1S2FPR1CYFIP1MAPKAPK3PDE4BSLC31A2SYKANKFY1RHOGABCF2MAP3K2SRGAP2BSERPINB9SLC35F6CARD8APBB1IPPLEKHJ1GBP2ETS2FCHO2PA2G4P4SDSARL4CPLBD1SEMA4DNME6CAB39PTPREDIAPH2-AS1CECR1ZNF571SNX10NCOA4SLAEMP3C2orf69TRAPPC2LCST3ENGFAM129AAPOC1DNAJC5CMTM7RELLIPADHODHSNCATFE3RHBDF2SNORA75TPD52L2ARHGAP18GKHEXBHPGDSMYO9BARL6IP4CCDC88APHLDA1JAZF1ZMIZ1LINC01094PLTPLCP2MAFBMKL1APH1BCYB5R4FAM49ARRM2BGDAP2FNIP2STARD5SLC37A2CASP1KLF4GNAI2NFKBIANXT2CCL2MFSD1MFSD2AAPOC2DAPP1CMKLR1CREBL2DERAIFI27L2HMGA1JUNBRGS10COTL1GNA13GAPTCSRNP1RASSF4CSF2RBSTARD3NLITGB3ELL2C3LOC102724297DAB2RAC2OTUD1PLA2G7TPP1SLC25A24SLC1A3CSF3RSELPLGRIPK2SLC29A1UCP2SAMD9LTNFRSF1BTICAM1LOC105371414CPED1GSKIPSTK40TREM1CAPGTINF2DOK3KLF2CD37PYCARDHCAR3PRDM1EMBLACTBCD4OLR1CCL8HCSTPLCG2RASSF5VMO1DOCK2HCKPYCARD-AS1RAB20CPVLPREX1SLAMF8VAV1KCTD12TMEM192BCL2L1STAT5ACCR1BLNKCIDEBMMDTTC38MAN2B1DPEP2RNVU1-20LRRC25FYBRAB10CD93HK3RNF149CARD16SIGLEC9LMO2FGD2PILRASP140WDR45BMS4A4AADAM28JAMLLINC01272IL6RCSD1NMBS100A9GPR183CXCL2ALCAMLILRB2LY96PECAM1FERMT3CD163NCF4NLRP3CSF1RAPCSTAB1MPP1SLC15A3MRC1TMEM52BGGTA1PGMFGEPB41L3EVI2BCD86FOLR2TBXAS1LAT2PALD1CCL20PLEKCLEC7ACKLFCXCR4TNFAIP3SPP1PTGS2LY86CXCL3MLKLFCGR3BCD84SCDFCGR1CPIFI30FCGR1BCYBAMGAT5IKZF1VASH1FCGR1ALPAR5RNASE6CD68SIGLEC14LGALS9FPR3SGK1CD53BMP2KMAP3K8ADAP2BCAT1ACSL1IQGAP2MAFPLA2G16CXCL16CTSSARRB2ITGAMFCER1GGPR132GIMAP4CCL3L3ARPC1BHAVCR2TREM2LYZLCP1AP1B1MPEG1CD74SRGNIL1BHLA-DQA1CD83CCL3L1C1orf162GPR65ITGAXRGS2ICAM1SLC7A7PTPRCC1QCPLAURFGL2C1QBALOX5APITGB2THEMIS2CCL4L1RNASET2IGSF21HBEGFCYBBFTH1P3CCL4L2CCL3CCL4NFATC2C3AR1C5AR1TNFSF13MS4A6AFCGR2BDOCK8PIK3CGTLR4NCF2FCGR3AIGSF6GPR34FCGR2CTYROBPMSR1FCGR2AHLA-DRB5CXCL8LAPTM5RGS1SLCO2B1HCLS1C1QACD14MNDAMS4A7RAPGEF2RBPJTSG101OPHN1NDUFAF1RPS15AP10EXOSC9C12orf10ZNF605PIGVRAB15RDH11NCOA1TMEM126BGINM1ANXA11CAP1COG3STK3ELF1ZNF438NBPF15SCARNA18YWHAEMYH9EIF2S3SLC44A1MED4KAT6ALSG1ORC4UBXN4CBWD3VPS13BRNASEKTHUMPD1RNF170PINK1-ASHCFC1R1RNF7ITM2BLOC101928137GAL3ST4CHCHD2NDUFA3RBM6RNPC3RNF130VHLSLC35F5NDUFA1SNAP23TSPAN31COPS5TBCBCCNINFKBIZZNF639TRA2AKIDINS220ETNK1CLCN3PTRFNBR1FCGRTHNRNPRTBC1D24HSPB1FIS1SDF2ZNF138HSPH1LGALSLSNX6TMBIM4TXNRD1KMT5ADYNLL1LAMTOR1PUM3BLOC1S6AHNAKSNAPC1METTL23PKMAPCDD1SRP9VPS8MORF4L1RARBCCDC25ZFYVE19FKBP7HPF1HYPKARPC4IFT52UGGT1NDUFV3N4BP2L2CAPZBMFN2TNFRSF21GXYLT1ATP6V1C1ITM2CWASHC2AUBE2Q2P1EMC10NUCB2PGBD1EPB41L2TMED4HGSNATFAM13CMRPL51C3orf14C6orf106MESTRB1CC1WDR45MIR3652ATP5F1OST4ARMCX3PDIA3MEF2CCCDC174CNPTPT1MAGT1SIPA1L1PTK2USP9XHSD17B7HDDC2PFDN4SERF2TSPAN6PWP1SLC50A1B4GALT3EIF5BTMEM45ATJP1ERLIN2PQLC3TNKSNADKNKAPUBL5HPRT1MED4-AS1VEGFBRAB22AINTS10CDH13CBWD6MAD2L1BPATP5JTSTA3LSM3ITFG2ANXA5PLIN2C6orf89EIF4G3MBD3EXOSC8TSR2NUTM2A-AS1LEPRCSPP1NBPF8ERGIC2RGS16CENPCNUDT10LOC101929340RGCCNUPL2SERP1HUWE1UBE2D4VRK3RBM3MGST3LYRM2RELL1ADAM9FAM8A1WTAPMAGOHBMETTL4RFLNBSUGT1IFT27TFAMEIF3IFKBP11METTL17LOC100419170EIF2S1NUTF2EIF3EFXYD6UBE4ATRIM22IFNGR1NDUFC2NORADPTBP2GPX4PRKD1FTOIRF7CDC26NMINAA20SBDSSYNPO2PRMT7CDH11COX4I1BET1LSSBEBLN3PHCFC2DNAJB4DPYSL2TRIM59UBE2FVIMACTN1STAT1PPP4R3BCRIPTSSR4MORC4TLE1DDB2TMBIM6COA1PURACYP51A1NOC3LSRPXRRAGBANAPC10RNF135CCDC51LYARANKHDTX4BRK1SLC35F1ARF5FSTL1PMEPA1MPDU1UFL1JAG1REEP3SUCLA2GLG1ZNF721LRRC57ITGB5CCDC82GUCD1ELOVL5ATOX1PDS5BPOLR2KPIP4K2CNNMTABHD18IGDCC4SBNO1TTC37GSNIDH3AITGBL1FASKLF10NAGKDNASE1L1RAB12SENP6KARSEIF4EGUSBSNORA104SUMO3PSMA1ODF2NPTN-IT1THAP4DNAJB11PHAXDIAPH2CNEP1R1HSPBAP1ANXA4CD160ARHGAP5TMEM59HSD17B10SPARCCOMTC3orf38SPCS3HNRNPA0ARHGAP17ZNF652MBNL1POLLTCEAL4SPC25SUMO2TSC22D1TMEM14AUSP2PIGCSMC6SLC2A3COPS8TOMM7DNAJC13ITGB1BP1FRY-AS1C4orf3OGTMAGOHMETAP2STRN3TMLHELOC102724552MYO19RNF111PPP1R12AOSR2ZC3H7AMAGI3PLNEMP2HIPK1SBDSP1AMPD2INTUANKRD10ZNF277DIDO1ITGB1RBMS3USP7FAM229BMPC1BANF1TAGLNTPBGSPOPFBXO7CTCFHNRNPUSGCERDXLINC01420LOC284023ZHX1DDX3YKIAA0040HNRNPH2OMA1ELK4TUBB6SPCS2SLC7A6FAM114A1HOTAIRALG13BEX3USP3SLC35B4NUB1ELOBDEGS1SDC2NT5CCHORDC1MAP4K4CIR1NBL1PPEF1COL4A1SAP30BPNBNIL1R1C1RSELENONKMT2CANXA7SPON2LAMTOR3SARSPDE5APTGR1FAM199XPTENMED12CDC42BPANDUFB10PLCE1ZNF451ERHUQCR11CMTM3SQSTM1ASH2LSAFBCSTF1MAPK1IP1LSMIM10L1SPIN1RWDD3LAMP2POGLUT1ZNF484NR1D2SNRPNSLC5A3RAB5BPSMD2ATP5C1KRBOX4KCTD20NREPC1QTNF5ZNF8THRAP3PSMC2STN1RAB40BARL2FRZBDDX39AFAHMYNNLOC105371814MNS1MLECPKN2SDHBRRBP1NDUFA13GTF2BTTLL3CCM2TBC1D15PMS2P3CTDSP1DBNDD2LYPLAL1SNU13USP31EIF5A2HIVEP3FAM111AZNF30TNFSF10SSSCA1TADA2BSNX2CRIP2THOC2CAMK1DHARSSMOC2GUCY1A3ATF6TSEN15CALRZNF333YIF1AFLRT2XAF1STX2LOC143666HAAOTCEAL9KCTD15MAP4DOCK1FAM168ACDK5SSPNB4GALT1SMAP1SSTR2RHOBTB3ZNF468SCML1ATG10SLC26A2HOOK3RHBDD3SGPP1CFHSRP54TMEM251LOC100131289GSTA4P4HA1FAM110BCD63RIN2DSELERVK3-1TRIM8WHAMMHDAC9CDKL1PSMB4CASC4RNF216A2MLEPROTL1ITPRIPL2TEX261MCM6DPYDDDR2UBE2D3URODPDZD11NBEAL1PSME1CAV1ARF6GBP1EIF2B3TOR1AIP1MIER1PSMA3PHLDB1SWI5GTF2E1TPM1RBM17USP22PARP8LIMS1TIMP2ZNF324AIDAOXCT1CD302RNASELFBXO42NEIL2ACVR1BABHD10NID1UBE2E2ARHGAP1STRAPZC4H2TBRG1AJUBAMRPS15EDEM1MCAMSLC41A2PDCD5BMP8ACALD1EFTUD2LIN37HDGFRP3IGFBP5PLS3MTERF4RRP1BCRTAPCALUITGAEMTCH1FKBP9SAMHD1STEAP2ADAMTS4BCAP29LTBP2TMEM212SLC6A6TRAPPC5SDF2L1PROS1GUCY1B3CYBRD1IFITM3ATL3PHTF1TAF8SPTBN1RAB3GAP2SARNPHLTF-AS1MRPL33MRPS18CESAMFKBP9P1APOL2SH3GLB1STX7DLGAP1-AS1MIR99AHGMPRIPRAD51BRAB21LOXSPRY4-IT1S100A6PARP14RPEACLYHDAC8C10orf10ARFGEF1GTF2H5ATF2DLDMRPL15AGPAT5PGM3TSC22D3C8orf44MALSU1TMEM30AGPC1EPS8CRY2KATNBL1PMP22DIS3FRMD6FRYACAD11TAF5LFMODFANCGSPG20KDELR3RBAKMKLN1-ASFGFR1OP2SLC12A8DOPEY1RRP15ZBTB38PSENENCDC37L1BTN2A1ZEB2INHBAPARP4VDAC3CMAHPLINC00963CMTR2ARL1KIAA1551JAK1ATRAIDZNF419MRPS11DPYSL3CYR61FIBING3BP2ROR2EHD2SEC24DERO1BCLEC2BALKBH5FBXL5CDC5LANO1MAN1A1CAPN2CAV2DOCK5SYNGR1XRRA1PARPBPSMC1ATDP2FAM208AANGPTL1C15orf39GNA12PON2CCZ1GLT8D2BTN3A2ZFP90FILIP1LFUT8CFHR1CCZ1BNCOR1MDP1CNIH4LINC00989RFTN2OR2A7GPX8MYO1BLXNZNF280DNDUFA6CPEWWC3FAM179BGLIPR2NAA10LOC101928093GLUD1OSMRUSP13TNS1FAM198BSTT3AOTUD6B-AS1TMX1DYNC1LI1DCBLD2PJA2TFB1MNLGN4YRNF11CTPS2MCFD2ZNF394SREK1IP1ARHGAP32DSTNIPAL2TMEM11SGMS2FBLN5OAFSNRNP35GJC1CLCN5LAMP5RASSF8ZNF684CHURC1SPATS2LFGD5-AS1COX17BROXSRGAP2DVAMP5PKD2LENG1TNFRSF12ACDK3IDSMFAP4PLAUA1BGPLXNC1EDNRAITGAVPRKCDBPNAP1L3PLAGL1LRCH3SERPINB6ANGPT2PDLIM3WNT5ATGFBIMPP5MRVI1HINT2YAE1D1PTPN12RITA1ZNF773PELI2PELOCBX6OSBPL9CEP112AP3B1C1SST7LEFEMP2OLFML1MRPL14CXCL12KANK2TNRC6BARHGAP29SYNJ2BP-COX16C20orf27FLJ20021DEF8SYNJ2BPTMEM248TMF1MIR5047ADAAPOL1SPHK1PATL1CADM1MAP1BGRPIGFBP3PLEKHO2PDE1ATIMM9SGIP1EML4C7orf26IRF2BP2NDUFA12LRRC32TBX15RAP1GDS1TRAPPC2BC3orf80COL11A1TWSG1APPL2TMEM200AGOLIM4MMP16KCNS3OLFM1MDFICCD248DHRS4GGNBP2SMARCA1MAPRE2CCDC80FAT4GPR176SEC22BATL1POMT1ABCC9PDE3AADCY3ARHGAP28MMP2ZBTB39EXT1PTGISGGT5CHST3ABCA9COL10A1EBF1STEAP1ALDH1A3IL32TIMM22PINK1LRIG2COPZ2DCAF5SQRDLTACC1LEPROTVGLL3THBS4TMEM35BFRKTMEM120APDGFRAAXLTRILHOPXTMEM204TIMP3FMO3IGFBP4GPR68PLA2G5SCN7APTNLOC100506388FAM200BABCA6UNC5BADAMTSL3MAFG-AS1RUNX1T1CD109LRP10RNF185OR51E1TMEM219LRRC15DIO2PTP4A3PEAR1LOC101927311HIGD1BALDH1L2TGFB3SGF29OCEL1LBHHMCN1PLPP1COL8A1EEA1EGLN3EPRSWISP1SYNE2CDK14SLC2A13MESDC2CD99TCF4FAM20CRAI2HEYLSGCBMORN2SELENOWFBN1CDH6FABP5COL5A1ANTXR2RPP30MRPL2L3MBTL3CRIP1PLXND1SEC23APPP6CCTHRC1ADGRF5ALDH1A1NAV1LOC400043VOPP1ARID5BPARP9RARRES2FHL1HEG1BICC1PKIGLRP1HSD17B11MINCRMCUBSH3KBP1GNG12NOTCH3ERAP2PRRG1ADIRFIFI16PLSCR4SPOCK1PLAC9IFITM1PRAF2ANTXR1ETV1SLC30A7SNED1FBLN1TGFBR2SEMA5AVSTM4NID2SYNE1-AS1GSTM3TP53INP1FAPOMDNF2SYT11THBS2DAD1S100A3TPPP3FN1ZNF829SERPING1PLXDC1SPARCL1EPAS1ZFHX4CAMK2N1MXRA5SERPINE1INPP4BCOL12A1DNAJC15APODDCNSFRP4PRRX1ITGA1CNRIP1CTSKCNN3SFRP2MIR4435-2HGMAP1AIGF1NDUFA4L2SRPX2RGS5MMP11COL5A2HTRA3TNFAIP6RAI14TWIST1ADAM19NRP1MXRA8LHFPLAMA4ZEB1COL18A1EEPD1NTMCRISPLD2GNG11IGFBP7COL6A2PRSS23POSTNAEBP1ISLRLUMPTGFRNECM2CYTORTFPIPDGFRBCOL6A1ID3PCOLCEC3orf70COL14A1BGNDLC1TIMP1ASPNCOL3A1SULF1COL1A1SLIT2COL1A2THY1COL6A34335443353433504334743165SNRNP40ADNPSNHG25CLDN4CDK6FAT1RPAINMTHFD2SHC4WBP1GOLPH3SNHG3SCUBE3MAEACOL27A1LINC01296CCNL2SMAD5FRMD4AC21orf91TGFBR3RARSEGFRKRT18SORBS2TCERG1FZD7NDUFA7MEIS1BOCFXYD3OBP2AOBP2BART3COL9A1SCHIP1SFRP1GTF2IP1WDR6ANXA3IGF1RCMYA5TBX3MGST1NUDT5TSTD1BCL11AGAS5RPL7AEFNA1SIK1PGAP1PHKG1CXADRSAA1RPS21SAA2SMIM4RPL11TTC19ARID1APARD3SMA4HP1BP3PCMTD1CCNT2MBNL2COL9A2GRHL1IRF6RSF1VCANDSG2KRT8SCNN1GFLNBCSTF3ISG20L2MYCPNISRFGFR1CRIM1SRPK2VITANKRD36KDSRMT2ARTF1ITGB4PIK3R3PRELPSOX11PRKCIFBLMAB21L3KANK4EHFPEG3SETD5PRKYPER2SERPINA3ITGA6NPIPB13ESRP1KLF5PIK3R1PPM1KFBXO32TCF7L1GPATCH8NPIPB11PARD6BRBPMSSNX22ERRFI1MPZL1NAV2ATP5A1USP34CRABP2KRT19PUM1GABRPPEG3-AS1CAPN12TM4SF1PADI2SCRG1AKAP9FABP7JUPATF4YAP1TFAP2CDSC2PAPSS1MZT2AMLXIPRNPS1MARCKSL1KRT15PYGBNRP2CTBP2H19NPIPB4NPIPB5TUFMPTPRFNIPAL3NPIPB3DSC3KRT23ACTG2FGFR2MIAFAM60AMALAT1APPCD24PDE9ADSPTRIM2AQP1NFIBEPCAMCDH1KRT14BAMBIKRT5IGF2

0 4 8 12

Value

04

00

00

Color Key

and Histogram

Count

A

B D

C

Fig 3


https://doi.org/10.1101/818260


PIK3CG

TLR4

NCF2

FCGR3A

IGSF6

GPR34

FCGR2C

TYROBP

MSR1

FCGR2A

HLA-DRB5

CXCL8

LAPTM5

RGS1

SLCO2B1

HCLS1

C1QA

CD14

MNDA

MS4A7

ECM2

CYTOR

TFPI

PDGFRB

COL6A1

ID3

PCOLCE

C3orf70

COL14A1

BGN

DLC1

TIMP1

ASPN

COL3A1

SULF1

COL1A1

SLIT2

COL1A2

THY1

COL6A3

DSC3

KRT23

ACTG2

FGFR2

MIA

FAM60A

MALAT1

APP

CD24

PDE9A

DSP

TRIM2

AQP1

NFIB

EPCAM

CDH1

KRT14

BAMBI

KRT5

IGF2

0

2

4

6

8

10

12

Value

010002000

Color Key

and Histogram

Count

Tum

or_

C02

Tum

or_

C05

Tum

or_

C06

Tum

or_

C07

Tum

or_

C08

Tum

or_

C14

Tum

or_

C15

Tum

or_

C16

Tum

or_

C20

Tum

or_

C22

Tum

or_

C23

Tum

or_

C24

Tum

or_

C27

Tum

or_

C28

Tum

or_

C32

Tum

or_

C33

Tum

or_

C37

Tum

or_

C38

Tum

or_

C40

Tum

or_

C43

Tum

or_

C49

Tum

or_

C50

Tum

or_

C51

Tum

or_

C57

Tum

or_

C58

Tum

or_

C60

Tum

or_

C62

Tum

or_

C63

Tum

or_

C66

Tum

or_

C68

Tum

or_

C70

Tum

or_

C71

Tum

or_

C72

Tum

or_

C73

Tum

or_

C74

Tum

or_

C76

Tum

or_

C77

Tum

or_

C79

Tum

or_

C81

Tum

or_

C82

Tum

or_

C84

Tum

or_

C85

Tum

or_

C86

Tum

or_

C87

Tum

or_

C88

Tum

or_

C89

Tum

or_

C90

Tum

or_

C92

Tum

or_

C93

Tum

or_

C94

Tum

or_

C95

Tum

or_

C01

Tum

or_

C04

Tum

or_

C26

Tum

or_

C41

Tum

or_

C42

Tum

or_

C53

Tum

or_

C69

Tum

or_

C10

Tum

or_

C12

Tum

or_

C19

Tum

or_

C34

Tum

or_

C75

Tum

or_

C91

PIK3CGTLR4NCF2FCGR3AIGSF6GPR34FCGR2CTYROBPMSR1FCGR2AHLA-DRB5CXCL8LAPTM5RGS1SLCO2B1HCLS1C1QACD14MNDAMS4A7ECM2CYTORTFPIPDGFRBCOL6A1ID3PCOLCEC3orf70COL14A1BGNDLC1TIMP1ASPNCOL3A1SULF1COL1A1SLIT2COL1A2THY1COL6A3DSC3KRT23ACTG2FGFR2MIAFAM60AMALAT1APPCD24PDE9ADSPTRIM2AQP1NFIBEPCAMCDH1KRT14BAMBIKRT5IGF2

0 2 4 6 8 10 12

Value

01

00

02

00

0

Color Key

and HistogramC

ount

Tum

or_

C02

Tum

or_

C05

Tum

or_

C06

Tum

or_

C07

Tum

or_

C08

Tum

or_

C14

Tum

or_

C15

Tum

or_

C16

Tum

or_

C20

Tum

or_

C22

Tum

or_

C23

Tum

or_

C24

Tum

or_

C27

Tum

or_

C28

Tum

or_

C32

Tum

or_

C33

Tum

or_

C37

Tum

or_

C38

Tum

or_

C40

Tum

or_

C43

Tum

or_

C49

Tum

or_

C50

Tum

or_

C51

Tum

or_

C57

Tum

or_

C58

Tum

or_

C60

Tum

or_

C62

Tum

or_

C63

Tum

or_

C66

Tum

or_

C68

Tum

or_

C70

Tum

or_

C71

Tum

or_

C72

Tum

or_

C73

Tum

or_

C74

Tum

or_

C76

Tum

or_

C77

Tum

or_

C79

Tum

or_

C81

Tum

or_

C82

Tum

or_

C84

Tum

or_

C85

Tum

or_

C86

Tum

or_

C87

Tum

or_

C88

Tum

or_

C89

Tum

or_

C90

Tum

or_

C92

Tum

or_

C93

Tum

or_

C94

Tum

or_

C95

Tum

or_

C01

Tum

or_

C04

Tum

or_

C26

Tum

or_

C41

Tum

or_

C42

Tum

or_

C53

Tum

or_

C69

Tum

or_

C10

Tum

or_

C12

Tum

or_

C19

Tum

or_

C34

Tum

or_

C75

Tum

or_

C91



Cluster_1

Cluster_2

Cluster_3

Ex

pre

ss

ion

(lo

g2

)

0

2

4

6

8

1 0

Ex

pre

ss

ion

(lo

g2

)

0

2

4

6

8

1 0

Ex

pre

ss

ion

(lo

g2

)

0

2

4

6

8

1 0

Ex

pre

ss

ion

(lo

g2

)

0

2

4

6

8

1 0

Ex

pre

ss

ion

(lo

g2

)

0

5

1 0

1 5

Ex

pre

ss

ion

(lo

g2

)

0

5

1 0

1 5

Ex

pre

ss

ion

(lo

g2

)

0

2

4

6

8

1 0

Ex

pre

ss

ion

(lo

g2

)

0

2

4

6

8

Ex

pre

ss

ion

(lo

g2

)

0

2

4

6

8

1 0

1 2

Ex

pre

ss

ion

(lo

g2

)

0

2

4

6

8

1 0

Ex

pre

ss

ion

(lo

g2

)

0

2

4

6

8

1 0

Ex

pre

ss

ion

(lo

g2

)

0

2

4

6

8

1 0

CK14

100× 200×

-30

0

30

-25 0 25 50

tsne1

tsne2

labels

Cluster_1

Cluster_2

Cluster_3

-30

0

30

-25 0 25 50

tsne1

tsne2

labels

Cluster_1

Cluster_2

Cluster_3

Cluster_1

Cluster_2

Cluster_3

A

B D

C

KRT5 KRT14 KRT23

EPCAM CDH1 CD24

COL1A2 THY1 BGN

CD14 CXCL8 MNDA

Fig 4


https://doi.org/10.1101/818260

NF

IBIG

F2

PD

E9A

MA

LA

T1

AC

TG

2C

D24

EP

CA

MB

AM

BI

TR

IM2

MIA

CD

H1

KR

T23

KR

T14

FA

M60A

KR

T5

DS

C3

AQ

P1

AP

PD

SP

FG

FR

2

NFIBIGF2PDE9AMALAT1ACTG2CD24EPCAMBAMBITRIM2MIACDH1KRT23KRT14FAM60AKRT5DSC3AQP1APPDSPFGFR2

-1 0 1

Value

03

0

Color Key

and Histogram

Count

MA

LA

T1

PD

E9A

NF

IBF

AM

60A

DS

P

TR

IM2

EP

CA

MC

DH

1B

AM

BI

MIA

AP

PA

QP

1IG

F2

AC

TG

2F

GF

R2

KR

T23

DS

C3

KR

T14

KR

T5

MALAT1

PDE9ANFIB

FAM60ADSP

TRIM2EPCAM

CDH1BAMBIMIA

APPAQP1

IGF2ACTG2

FGFR2KRT23DSC3

KRT14KRT5

-1 0 1

Value

04

0

Color Key

and Histogram

Count

A

B

C

Cluster_1

Cluster_2

Cluster_3

CTC-candidate

CTC-confirmed

Primary tumor

PCa cell line

White blood cell

Tu

mo

r_C

02

Tu

mo

r_C

05

Tu

mo

r_C

06

Tu

mo

r_C

07

Tu

mo

r_C

08

Tu

mo

r_C

14

Tu

mo

r_C

15

Tu

mo

r_C

16

Tu

mo

r_C

20

Tu

mo

r_C

22

Tu

mo

r_C

23

Tu

mo

r_C

24

Tu

mo

r_C

27

Tu

mo

r_C

28

Tu

mo

r_C

32

Tu

mo

r_C

33

Tu

mo

r_C

37

Tu

mo

r_C

38

Tu

mo

r_C

40

Tu

mo

r_C

43

Tu

mo

r_C

49

Tu

mo

r_C

50

Tu

mo

r_C

51

Tu

mo

r_C

57

Tu

mo

r_C

58

Tu

mo

r_C

60

Tu

mo

r_C

62

Tu

mo

r_C

63

Tu

mo

r_C

66

Tu

mo

r_C

68

Tu

mo

r_C

70

Tu

mo

r_C

71

Tu

mo

r_C

72

Tu

mo

r_C

73

Tu

mo

r_C

74

Tu

mo

r_C

76

Tu

mo

r_C

77

Tu

mo

r_C

79

Tu

mo

r_C

81

Tu

mo

r_C

82

Tu

mo

r_C

84

Tu

mo

r_C

85

Tu

mo

r_C

86

Tu

mo

r_C

87

Tu

mo

r_C

88

Tu

mo

r_C

89

Tu

mo

r_C

90

Tu

mo

r_C

92

Tu

mo

r_C

93

Tu

mo

r_C

94

Tu

mo

r_C

95

Tu

mo

r_C

01

Tu

mo

r_C

04

Tu

mo

r_C

26

Tu

mo

r_C

41

Tu

mo

r_C

42

Tu

mo

r_C

53

Tu

mo

r_C

69

Tu

mo

r_C

10

Tu

mo

r_C

12

Tu

mo

r_C

19

Tu

mo

r_C

34

Tu

mo

r_C

75

Tu

mo

r_C

91

Pr1

0.1

.2P

r10

.1.3

Pr1

0.2

.1P

r11

.1.2

Pr1

1.2

.1P

r11

.2.3

Pr1

2.1

.1P

r12

.1.2

Pr1

3.1

.1P

r13

.1.2

Pr1

4.3

.2P

r14

.3.3

Pr1

5.1

.2P

r16

.1.1

Pr1

6.1

.2P

r16

.1.3

Pr1

6.1

.4P

r16

.1.6

Pr1

8.1

.2P

r18

.1.3

Pr1

8.1

.9P

r18

.2.4

Pr2

0.1

.2P

r22

.1.1

Pr2

2.1

.3P

r9.1

.5P

r9.1

.7P

r9.2

.1P

r9.2

.2P

r9.3

.2P

r9.3

.3P

r9.3

.4P

r9.3

.5P

r1.1

.1P

r2.1

.1P

r2.1

.2P

r3.1

.2P

r4.1

.1P

r4.1

.2P

r5.1

.1P

r5.1

.2P

r5.1

.3P

r6.1

.3P

r6.1

.4P

r8.1

.2P

r10

.2.2

Pr1

1.1

.1P

r11

.2.2

Pr1

1.3

.1P

r11

.3.2

Pr1

1.3

.3P

r11

.4.1

Pr1

1.4

.2P

r11

.4.3

Pr1

1.4

.4P

r11

.4.5

Pr1

1.4

.6P

r14

.1.1

Pr1

4.2

.1P

r14

.2.2

Pr1

4.2

.3P

r14

.2.4

Pr1

4.2

.5P

r14

.2.6

Pr1

4.3

.1P

r14

.3.4

Pr1

4.3

.5P

r14

.3.6

Pr1

5.1

.1P

r17

.1.1

Pr1

7.1

.2P

r17

.2.1

Pr1

7.2

.2P

r9.1

.1P

r9.1

.2P

r9.1

.3P

r9.1

.4P

r9.1

.6P

r9.3

.1P

r9.3

.6P

r9.3

.7P

r9.3

.8P

r18

.1.1

Pr1

8.1

.4P

r18

.1.5

Pr1

8.1

.6P

r18

.1.7

Pr1

8.1

.8P

r18

.2.1

Pr1

8.2

.2P

r18

.2.3

Pr1

9.1

.2P

r19

.1.3

Pr1

9.1

.4P

r19

.1.5

Pr2

0.1

.1P

r21

.1.1

Pr2

1.1

.10

Pr2

1.1

.11

Pr2

1.1

.12

Pr2

1.1

.2P

r21

.1.3

Pr2

1.1

.4P

r21

.1.5

Pr2

1.1

.6P

r21

.1.7

Pr2

1.1

.8P

r21

.1.9

Pr2

2.1

.10

Pr2

2.1

.11

Pr2

2.1

.12

Pr2

2.1

.2P

r22

.1.4

Pr2

2.1

.5P

r22

.1.6

Pr2

2.1

.7P

r22

.1.8

Pr2

2.1

.9P

r3.1

.1P

r6.1

.1P

r6.1

.2P

r6.1

.5P

riT

um

1P

riT

um

2P

riT

um

3P

riT

um

4P

riT

um

10

Pri

Tu

m1

1P

riT

um

12

Pri

Tu

m5

Pri

Tu

m6

Pri

Tu

m7

Pri

Tu

m8

Pri

Tu

m9

DU

14

5.1

DU

14

5.2

DU

14

5.3

DU

14

5.4

DU

14

5.5

LN

Ca

P.1

LN

Ca

P.2

LN

Ca

P.3

LN

Ca

P.4

LN

Ca

P.5

LN

Ca

P.D

.1L

NC

aP

.D.2

LN

Ca

P.D

.3L

NC

aP

.D.4

LN

Ca

P.D

.5L

NC

aP

.R.1

LN

Ca

P.R

.2L

NC

aP

.R.3

LN

Ca

P.R

.4L

NC

aP

.R.5

PC

3.1

PC

3.2

PC

3.3

PC

3.4

PC

3.5

VC

aP

.1V

Ca

P.2

VC

aP

.3V

Ca

P.4

VC

aP

.5P

r23

.1.w

1P

r23

.1.w

2H

D1

.1.w

1H

D1

.1.w

2H

D1

.1.w

3

AQP1

NFIB

EPCAM

CDH1

KRT14

BAMBI

KRT5

IGF2

0 1 2 3

Value

0

Color Key

and Histogram

Count

SC

_90

57

SC

_90

22

MO

_1

07

1T

P_2

03

4S

C_

90

62

SC

_90

01

MO

_1

09

4X

6115237

MO

_1

11

8M

O_1

21

5T

P_2

06

1X

6115242

SC

_90

38

MO

_1

05

4S

C_

90

12

MO

_1

02

0S

C_

90

31

X6115122

SC

_90

07

TP

_2

00

9S

C_

90

37

SC

_90

60

SC

_90

48

SC

_90

17

SC

_90

94

MO

_1

17

9S

C_

90

34

TP

_2

02

0S

C_

90

71

MO

_1

33

6S

C_

90

46

MO

_1

24

9S

C_

90

43

SC

_90

23

SC

_90

49

SC

_90

61

X1115183

SC

_90

86

MO

_1

22

1M

O_1

23

2X

6115118

X1115202

X1115153

SC

_90

19

MO

_1

16

1M

O_1

08

4M

O_1

21

9S

C_

90

08

X6115227

SC

_90

92

MO

_1

19

2M

O_1

12

8X

1115157

SC

_90

80

TP

_2

03

2S

C_

90

58

X6115234

SC

_90

81

TP

_2

06

4X

1115161

SC

_90

28

MO

_1

33

9M

O_1

33

7M

O_1

11

4X

1115156

MO

_1

27

7S

C_

90

55

MO

_1

26

2M

O_1

31

6M

O_1

17

6M

O_1

18

4M

O_1

20

2X

1115244

SC

_90

93

SC

_90

83

X6115117

TP

_2

05

4S

C_

90

47

SC

_90

91

X6115115

MO

_1

24

4S

C_

90

30

SC

_90

59

MO

_1

09

5X

6115121

SC

_90

99

X6115114

SC

_90

16

SC

_90

72

SC

_90

29

X6115250

SC

_90

32

SC

_90

97

SC

_90

73

X6115233

SC

_90

63

MO

_1

12

4M

O_1

24

1X

6115224

X1115154

SC

_90

36

X6115251

SC

_90

26

SC

_90

18

X6115247

X6115219

SC

_90

09

TP

_2

00

1S

C_

90

54

TP

_2

01

0S

C_

90

50

X6115123

MO

_1

01

4M

O_1

04

0M

O_1

01

3T

P_2

06

0S

C_

90

10

SC

_90

68

APP

MALAT1

CDH1EPCAM

KRT5

KRT14DSC3

KRT23

FGFR2ACTG2

TRIM2BAMBI

IGF2

MIAAQP1

PDE9A

DSPNFIB

FAM60A

0 2 4 6 8 10

Value

03

00

Color Key

and Histogram

Count

Tum

or_

C02

Tum

or_

C05

Tum

or_

C06

Tum

or_

C07

Tum

or_

C08

Tum

or_

C14

Tum

or_

C15

Tum

or_

C16

Tum

or_

C20

Tum

or_

C22

Tum

or_

C23

Tum

or_

C24

Tum

or_

C27

Tum

or_

C28

Tum

or_

C32

Tum

or_

C33

Tum

or_

C37

Tum

or_

C38

Tum

or_

C40

Tum

or_

C43

Tum

or_

C49

Tum

or_

C50

Tum

or_

C51

Tum

or_

C57

Tum

or_

C58

Tum

or_

C60

Tum

or_

C62

Tum

or_

C63

Tum

or_

C66

Tum

or_

C68

Tum

or_

C70

Tum

or_

C71

Tum

or_

C72

Tum

or_

C73

Tum

or_

C74

Tum

or_

C76

Tum

or_

C77

Tum

or_

C79

Tum

or_

C81

Tum

or_

C82

Tum

or_

C84

Tum

or_

C85

Tum

or_

C86

Tum

or_

C87

Tum

or_

C88

Tum

or_

C89

Tum

or_

C90

Tum

or_

C92

Tum

or_

C93

Tum

or_

C94

Tum

or_

C95

Tum

or_

C01

Tum

or_

C04

Tum

or_

C26

Tum

or_

C41

Tum

or_

C42

Tum

or_

C53

Tum

or_

C69

Tum

or_

C10

Tum

or_

C12

Tum

or_

C19

Tum

or_

C34

Tum

or_

C75

Tum

or_

C91

Tumor_C15

Tumor_C14

Tumor_C08

Tumor_C07

Tumor_C06

Tumor_C05

Tumor_C02


Benign CD49f Hi

Benign CD49f Lo

Cancer CD49f Hi

Cancer CD49f Lo

BS

2771_C

ancerL

o

BS

2980_B

enig

nLo

BS

2980_C

ancerL

o

BS

2916_C

ancerL

o

BS

3337_C

ancerL

o

BS

3337_B

enig

nLo

BS

2506_B

enig

nLo

BS

2340_B

enig

nLo

BS

2339_B

enig

nLo

BS

2553_C

ancerL

o

BS

2553_C

ancerH

i

BS

2340_B

enig

nH

i

BS

2340_C

ancerH

i

BS

2506_B

enig

nH

i

BS

2339_C

ancerH

i

BS

2339_B

enig

nH

i

BS

2980_B

enig

nH

i

BS

2980_C

ancerH

i

BS

3337_B

enig

nH

i

BS

3337_C

ancerH

i

ACTG2

MIA

AQP1

IGF2

PDE9A

BAMBI

MALAT1

DSP

APP

CD24

CDH1

KRT14

FGFR2

KRT5

DSC3

FAM60A

TRIM2

KRT23

NFIB

EPCAM

0 5 15

Value

04

0

Color Key

and Histogram

Count

Tu

mo

r_C

10

Tu

mo

r_C

75

Tu

mo

r_C

91

Tu

mo

r_C

34

Tu

mo

r_C

19

Pr8

.1.2

Pr9

.1.7

Pr5

.1.3

Pr5

.1.2

Pr5

.1.1

Tu

mo

r_C

12

Tu

mo

r_C

66

Pr3

.1.2

HD

1.1

.w3

Pr1

1.2

.3P

r15

.1.2

HD

1.1

.w2

Pr1

8.1

.3P

r23

.1.w

1P

r14

.3.2

Pr9

.3.4

Pr2

.1.1

Pr1

3.1

.1P

r11

.4.3

HD

1.1

.w1

Pr1

2.1

.2P

r23

.1.w

2P

r16

.1.1

Pr6

.1.4

Pr9

.3.5

Pr1

2.1

.1P

riT

um

7P

riT

um

12

Pri

Tu

m6

Pri

Tu

m5

Tu

mo

r_C

33

Pri

Tu

m1

0P

riT

um

3P

riT

um

4P

riT

um

2P

riT

um

8P

riT

um

1P

riT

um

11

Pri

Tu

m9

Tu

mo

r_C

42

Tu

mo

r_C

04

Tu

mo

r_C

53

Tu

mo

r_C

41

Tu

mo

r_C

01

Tu

mo

r_C

69

Tu

mo

r_C

26

Tu

mo

r_C

43

Tu

mo

r_C

72

Tu

mo

r_C

58

Tu

mo

r_C

49

Tu

mo

r_C

15

Tu

mo

r_C

08

Tu

mo

r_C

79

Tu

mo

r_C

76

Tu

mo

r_C

94

Tu

mo

r_C

14

Tu

mo

r_C

05

Tu

mo

r_C

68

Tu

mo

r_C

37

Tu

mo

r_C

07

Tu

mo

r_C

23

Tu

mo

r_C

71

Tu

mo

r_C

82

Tu

mo

r_C

81

Tu

mo

r_C

28

Tu

mo

r_C

32

Tu

mo

r_C

06

Tu

mo

r_C

38

Tu

mo

r_C

16

Tu

mo

r_C

90

Tu

mo

r_C

73

Tu

mo

r_C

50

Tu

mo

r_C

92

Tu

mo

r_C

89

Tu

mo

r_C

63

Tu

mo

r_C

02

Tu

mo

r_C

60

Tu

mo

r_C

57

Tu

mo

r_C

85

Tu

mo

r_C

77

Tu

mo

r_C

27

Tu

mo

r_C

95

Tu

mo

r_C

22

Tu

mo

r_C

62

Tu

mo

r_C

24

Tu

mo

r_C

20

Tu

mo

r_C

51

Tu

mo

r_C

40

Tu

mo

r_C

84

Tu

mo

r_C

88

Tu

mo

r_C

70

Tu

mo

r_C

93

Tu

mo

r_C

74

Tu

mo

r_C

87

Tu

mo

r_C

86

Pr1

4.1

.1P

r10

.1.3

Pr1

6.1

.3P

r14

.3.4

Pr1

6.1

.2P

r10

.2.1

Pr1

3.1

.2P

r22

.1.1

2P

r18

.1.9

Pr1

0.2

.2P

r22

.1.1

Pr9

.3.3

Pr1

.1.1

Pr4

.1.1

Pr9

.3.6

Pr1

4.3

.5P

r14

.3.3

Pr1

1.1

.2P

r11

.1.1

Pr3

.1.1

Pr1

8.1

.5P

r22

.1.7

Pr1

7.1

.1P

r9.2

.2P

r16

.1.6

Pr1

4.2

.2P

r10

.1.2

Pr1

1.4

.4P

r2.1

.2P

r11

.2.2

Pr1

1.2

.1D

U1

45

.4P

r19

.1.3

Pr1

8.2

.4P

r11

.4.6

Pr1

9.1

.5P

r22

.1.1

0P

r14

.3.1

Pr1

1.4

.2P

r9.1

.6P

r19

.1.4

Pr1

8.1

.6P

r18

.2.3

Pr2

2.1

.9P

r14

.2.4

Pr2

2.1

.11

Pr9

.1.5

Pr2

1.1

.3P

r9.3

.7P

r21

.1.5

Pr2

1.1

.6P

r20

.1.2

Pr1

5.1

.1P

r16

.1.4

Pr2

2.1

.3P

r6.1

.3P

r18

.1.2

Pr9

.2.1

PC

3.5

PC

3.1

PC

3.4

PC

3.3

Pr1

8.2

.1P

r17

.2.2

Pr1

9.1

.2P

r18

.1.1

Pr2

1.1

.4P

r11

.3.1

LN

Ca

P.2

DU

14

5.5

LN

Ca

P.1

LN

Ca

P.R

.4L

NC

aP

.R.1

LN

Ca

P.3

LN

Ca

P.4

LN

Ca

P.5

LN

Ca

P.R

.2L

NC

aP

.D.3

LN

Ca

P.R

.3L

NC

aP

.R.5

LN

Ca

P.D

.1L

NC

aP

.D.2

LN

Ca

P.D

.5L

NC

aP

.D.4

Pr1

1.4

.5P

r18

.2.2

Pr2

1.1

.12

Pr1

4.2

.3P

r18

.1.7

Pr2

2.1

.2P

r14

.3.6

Pr1

4.2

.6P

r14

.2.5

Pr2

1.1

.11

Pr2

1.1

.1P

r21

.1.1

0P

r21

.1.9

Pr1

1.4

.1P

r21

.1.7

Pr2

1.1

.2P

r9.1

.1P

r22

.1.4

Pr9

.3.2

Pr9

.1.2

PC

3.2

DU

14

5.2

Pr9

.1.3

Pr9

.3.1

Pr6

.1.5

Pr2

2.1

.5P

r4.1

.2P

r20

.1.1

DU

14

5.3

DU

14

5.1

VC

aP

.3P

r14

.2.1

VC

aP

.5V

Ca

P.2

Pr1

1.3

.2V

Ca

P.1

VC

aP

.4P

r21

.1.8

Pr1

8.1

.8P

r17

.1.2

Pr2

2.1

.8P

r17

.2.1

Pr1

1.3

.3P

r6.1

.1P

r18

.1.4

Pr2

2.1

.6P

r9.3

.8P

r6.1

.2P

r9.1

.4


0 5 10 15

Value

04

00

08

00

0

Color Key

and Histogram

Count

Tu

mo

r_C

10

Tu

mo

r_C

75

Tu

mo

r_C

91

Tu

mo

r_C

34

Tu

mo

r_C

19

Pr8

.1.2

Pr9

.1.7

Pr5

.1.3

Pr5

.1.2

Pr5

.1.1

Tu

mo

r_C

12

Tu

mo

r_C

66

Pr3

.1.2

HD

1.1

.w3

Pr1

1.2

.3P

r15

.1.2

HD

1.1

.w2

Pr1

8.1

.3P

r23

.1.w

1P

r14

.3.2

Pr9

.3.4

Pr2

.1.1

Pr1

3.1

.1P

r11

.4.3

HD

1.1

.w1

Pr1

2.1

.2P

r23

.1.w

2P

r16

.1.1

Pr6

.1.4

Pr9

.3.5

Pr1

2.1

.1P

riT

um

7P

riT

um

12

Pri

Tu

m6

Pri

Tu

m5

Tu

mo

r_C

33

Pri

Tu

m1

0P

riT

um

3P

riT

um

4P

riT

um

2P

riT

um

8P

riT

um

1P

riT

um

11

Pri

Tu

m9

Tu

mo

r_C

42

Tu

mo

r_C

04

Tu

mo

r_C

53

Tu

mo

r_C

41

Tu

mo

r_C

01

Tu

mo

r_C

69

Tu

mo

r_C

26

Tu

mo

r_C

43

Tu

mo

r_C

72

Tu

mo

r_C

58

Tu

mo

r_C

49

Tu

mo

r_C

15

Tu

mo

r_C

08

Tu

mo

r_C

79

Tu

mo

r_C

76

Tu

mo

r_C

94

Tu

mo

r_C

14

Tu

mo

r_C

05

Tu

mo

r_C

68

Tu

mo

r_C

37

Tu

mo

r_C

07

Tu

mo

r_C

23

Tu

mo

r_C

71

Tu

mo

r_C

82

Tu

mo

r_C

81

Tu

mo

r_C

28

Tu

mo

r_C

32

Tu

mo

r_C

06

Tu

mo

r_C

38

Tu

mo

r_C

16

Tu

mo

r_C

90

Tu

mo

r_C

73

Tu

mo

r_C

50

Tu

mo

r_C

92

Tu

mo

r_C

89

Tu

mo

r_C

63

Tu

mo

r_C

02

Tu

mo

r_C

60

Tu

mo

r_C

57

Tu

mo

r_C

85

Tu

mo

r_C

77

Tu

mo

r_C

27

Tu

mo

r_C

95

Tu

mo

r_C

22

Tu

mo

r_C

62

Tu

mo

r_C

24

Tu

mo

r_C

20

Tu

mo

r_C

51

Tu

mo

r_C

40

Tu

mo

r_C

84

Tu

mo

r_C

88

Tu

mo

r_C

70

Tu

mo

r_C

93

Tu

mo

r_C

74

Tu

mo

r_C

87

Tu

mo

r_C

86

Pr1

4.1

.1P

r10

.1.3

Pr1

6.1

.3P

r14

.3.4

Pr1

6.1

.2P

r10

.2.1

Pr1

3.1

.2P

r22

.1.1

2P

r18

.1.9

Pr1

0.2

.2P

r22

.1.1

Pr9

.3.3

Pr1

.1.1

Pr4

.1.1

Pr9

.3.6

Pr1

4.3

.5P

r14

.3.3

Pr1

1.1

.2P

r11

.1.1

Pr3

.1.1

Pr1

8.1

.5P

r22

.1.7

Pr1

7.1

.1P

r9.2

.2P

r16

.1.6

Pr1

4.2

.2P

r10

.1.2

Pr1

1.4

.4P

r2.1

.2P

r11

.2.2

Pr1

1.2

.1D

U1

45

.4P

r19

.1.3

Pr1

8.2

.4P

r11

.4.6

Pr1

9.1

.5P

r22

.1.1

0P

r14

.3.1

Pr1

1.4

.2P

r9.1

.6P

r19

.1.4

Pr1

8.1

.6P

r18

.2.3

Pr2

2.1

.9P

r14

.2.4

Pr2

2.1

.11

Pr9

.1.5

Pr2

1.1

.3P

r9.3

.7P

r21

.1.5

Pr2

1.1

.6P

r20

.1.2

Pr1

5.1

.1P

r16

.1.4

Pr2

2.1

.3P

r6.1

.3P

r18

.1.2

Pr9

.2.1

PC

3.5

PC

3.1

PC

3.4

PC

3.3

Pr1

8.2

.1P

r17

.2.2

Pr1

9.1

.2P

r18

.1.1

Pr2

1.1

.4P

r11

.3.1

LN

Ca

P.2

DU

14

5.5

LN

Ca

P.1

LN

Ca

P.R

.4L

NC

aP

.R.1

LN

Ca

P.3

LN

Ca

P.4

LN

Ca

P.5

LN

Ca

P.R

.2L

NC

aP

.D.3

LN

Ca

P.R

.3L

NC

aP

.R.5

LN

Ca

P.D

.1L

NC

aP

.D.2

LN

Ca

P.D

.5L

NC

aP

.D.4

Pr1

1.4

.5P

r18

.2.2

Pr2

1.1

.12

Pr1

4.2

.3P

r18

.1.7

Pr2

2.1

.2P

r14

.3.6

Pr1

4.2

.6P

r14

.2.5

Pr2

1.1

.11

Pr2

1.1

.1P

r21

.1.1

0P

r21

.1.9

Pr1

1.4

.1P

r21

.1.7

Pr2

1.1

.2P

r9.1

.1P

r22

.1.4

Pr9

.3.2

Pr9

.1.2

PC

3.2

DU

14

5.2

Pr9

.1.3

Pr9

.3.1

Pr6

.1.5

Pr2

2.1

.5P

r4.1

.2P

r20

.1.1

DU

14

5.3

DU

14

5.1

VC

aP

.3P

r14

.2.1

VC

aP

.5V

Ca

P.2

Pr1

1.3

.2V

Ca

P.1

VC

aP

.4P

r21

.1.8

Pr1

8.1

.8P

r17

.1.2

Pr2

2.1

.8P

r17

.2.1

Pr1

1.3

.3P

r6.1

.1P

r18

.1.4

Pr2

2.1

.6P

r9.3

.8P

r6.1

.2P

r9.1

.4


0 5 10 15

Value

0

Color Key

and Histogram

Count

Fig 5


https://doi.org/10.1101/818260

Single-cell dissection of a rare human prostate …specimens collected, after dissociation of the...

Documents

Transcript of Single-cell dissection of a rare human prostate …specimens collected, after dissociation of the...