Poster Department Presenting Designation Abstract Title...

Poster No.

Department Presenting Author

Designation Abstract Title Authors

Phd-13 Biochemistry Ms. Gunjan Sharma

PhD Student ETV6 as a regulator of the IGF2BP family of RNA binding proteins in Acute Lymphoblastic Leukemia

Gunjan Sharma, Elza Boby, Ayushi Jain, Parthaprasad Chattopadhyay, Anita Chopra, Sameer Bakhshi, Jayanth Kumar Palanichamy

Phd-14 Biochemistry Nitesh Mishra PhD Student Viral characteristics associated with maintenance of elite neutralizing activity in chronically HIV-1C infected monozygotic pediatric twins

Mishra Nitesh, Makhdoomi Ashraf Muzamil, Sharma Shaifali, Kumar Sanjeev, Kumar Deepshikha, Chawla Himanshi, Singh Ravinder, Kanga Uma, Das Kumar Bimal, Lodha Rakesh, Kabra Kumar Sushil, Luthra Kalpana

Phd-15 Biochemistry Amar Preet Kaur

PhD Student Evaluating the role of Metformin on differentiation and tumor microenvironment in human colorectal carcinoma

Kaur, Preet Amar, Dey, Devanjan, Bhattacharya, Aditi, Bansal, Aakriti, Das, Prasenjit, Vishnubhatla, Sreenivas, Sen, Sudip

Phd-16 Biochemistry Deepti Pathak PhD Student The Involvement of Iron in Neurodegeneration associated with Friedreich’s ataxia

Deepti Pathak, Achal Kumar Srivastava, Sheffali Gulati, M.V. Padma, Moganty R Rajeswari

Phd-17 Biochemistry Dr. Srinivas H PhD Student Characterisation of Fat1 processing and its interaction with β-catenin in glioma

Gowda H Srinivas, Gupta Yakhlesh, Malik Nargis, Goswami Sanjeev, Chattopadhyay Parthaprasad, Sarkar Chitra, Suri Ashish, Gupta K Deepak, Sinha Subrata, Chosdol Kunzang.

Phd-18 Biochemistry Vikas Kumar PhD Student Transactivation of HnRNPD/Auf-1 by NF-κB(P65/RelA) in oral cancer

Kumar Vikas, Kumar Manish, Chauhan S Shyam

Phd-21 Biochemistry Sudhasini Panda

PhD Student Role of vitamin D and its associated molecules in innate immunity modulation in pulmonary tuberculosis

Sudhasini Panda, Ambrish Tiwari, Kalpana Luthra, SK Sharma, Archana Singh

Phd-22 Biochemistry Nidhi Gupta PhD Student Targeting of verrsican by microRNAs inhibit multiple myeloma progression: A novel potential therapeutic strategy

Gupta Nidhi, Kumar Raman Seth Tulika, Garg Bhavuk, Sharma Alpana

Phd-28 Biochemistry Nargis Malik Phd. Student

FAT1 gene regulates

Programmed cell death

10 (PDCD10) via miR-

221 in glioblastoma

Nargis Malik,

Parthaprasad

Chattopadhyay,

Chitra Sarkar,

Ashish Suri,

Subrata Sinha,

Kunzang Chosdol

Phd-82 Biochemistry Khushwant Singh

PhD Student Silver nanoparticle enhance tumor associated macrophage function and decreases tumor cell proliferation

Gautam Kumar Pramod, Singh Khushwant, Anita, Garg Drishti, Bukhari Jehangir.

Phd-84 Biochemistry Himani Thakkar

PhD Student HDL abnormalities in cardiovascular disease: Quality or Quantity?

Thakkar Himani, Vincent Vinnyfred, Roy Ambuj, Singh Sandeep, Ramakrishnan Lakshmy, Singh Archna

Phd-89 Biochemistry Dayasagar Das

PhD Student Unraveling the role of Gamma Delta T cell subsets: Key players in the immunopathogenesis of Pemphigus Vulgaris

Das Dayasagar,KV Santosh, Arava Sudheer, Khandpur Sujay, Sharma Alpana

Phd-98 Biochemistry Mohit Arora PhD Student Elevated expression of Dipeptidyl Peptidase III correlates with the Tumor Infiltrating Lymphocyte(TILs) composition in multiple cancers.

Arora Mohit, Gupta Jasmeen, Prajapati Chand Subhash, Chauhan Singh Shyam

Phd-103

Biochemistry Charandeep Kaur

PhD Student Acute Febrile Illness associated malaria co-infections in tropical diseases: A spectrum and incidence in Indian population

Atreyi Pramanik, Charandeep Kaur, Rajendra Mandage, Vinod Kumar, Shounak Saha, Adarsh Singh, Manish Soneja, Pragyan Acharya

Phd-104

Biochemistry Yakhlesh Gupta

PhD Student FAT1, a novel regulator of YAP1- effector molecule of Salvador-Warts-Hippo (SWH) pathway, in human Glioblastoma

Gupta Yakhlesh, Shivajirao Sachin Shelke, Irshad Khushboo, Dikshit Bhawana, Srivastav Tapasya, Chattopadhyay Parthaprasad, Sinha Subrata & Chosdol Kunzang

Phd-109

Biochemistry Sanjeev Goswami

PhD Student Role of IGF2BP3 in migration and invasion of epithelial cancer cell lines

Sanjeev Goswami, Gunjan Sharma, Elza Boby, Parthaprasad Chattopadhyay, Jayanth Kumar Palanichamy

Phd-110

Biochemistry Mohamad Tarik

PhD Student Feasibility of measuring sodium, potassium and creatinine from urine sample on dried filter paper

Mohamad Tarik, Lakshmy Ramakrishnan, Ritvik Amarchand, Harshal R. Salve, Prashant Mathur, Pradeep Joshi, Anand Krishnan

Phd-125

Biochemistry Qazi Sahar PhD Student BESFA: Bioinformatics based Evolutionary, Structural & Functional Analysis In-silico analysis of POTE Paralogs

Qazi Sahar, Sharma Neetu, Karpf R Adam, Sharma Ashok

Phd-126

Biochemistry Nisha Manav PhD Student Epigenetic Regulation of Pericentromerically localized Cancer-Testis/Germline Antigen POTE and its use as diagnostic marker/ immunotherapeutic agent in Ovarian Cancer

Manav Nisha, Kundu Subhadip, Jha Sangit, Quazi Sahar, Sharma Neetu, Kumar Pawan, Vanamail P, Mathur Sandeep, Sharma J.B, Chauhan S.S, Kumar Lalit, Kumar Sunesh, Karpf R Adam,

Sharma Ashok

Phd-134

Biochemistry Aditi Bhattacharya

PhD Student Identifying molecules to distinguish between colorectal cancer stem cells and normal colonic stem cells to develop novel diagnostic and prognostic markers

Bhattacharya Aditi, Manhas Janvie, Dey Devanjan, Bhat Muzaffar, Das Prasenjit, Pal Sujoy, Deo SVS, Parshad Rajinder, Ghosh Debabrata, Sen Sudip

Phd-141

Biochemistry Devanjan Dey PhD Student Using human fetal neural stem cells and oligodendrocytes as a disease model to delineate the pathogenesis of cerebral palsy.

Dey D, Shrivastav V, Bhat MA, Singal C.M.S., Sharma JB, Palanichamy JK, Chattopadhyay P, Sinha S, Seth P, Sen S

Phd-13

Title: ETV6as a regulator of the IGF2BP family of RNA binding proteins in Acute

Lymphoblastic Leukemia

Authors:Gunjan Sharma1, Elza Boby

1, Ayushi Jain

1, Parthaprasad Chattopadhyay

1, Anita Chopra

3,

Sameer Bakhshi2, JayanthKumar Palanichamy

1

Affiliation and Address: 1Dept. of Biochemistry,

2Dept. of Medical Oncology,

3Dept. of Lab Oncology,

All India Institute of Medical Sciences, New Delhi, India

Email: [email protected]

Background: Acute lymphoblastic leukemia (ALL) is the most common childhood cancer,

with 85% of B-cell origin.ETV6-RUNX1 translocation is the most common genetic

aberrationin ALL. Deletion of the non-rearranged ETV6 allele is also seen commonly in

these patients. Insulin like Growth Factor 2 mRNA Binding Proteins, IGF2BP1 and

IGF2BP3are found to be specifically overexpressed inB-ALL patients with particular

translocations.We have tried to characterize the regulation IGF2BP1 and IGF2BP3 by ETV6

in ETV6-RUNX1 positive B-ALL.

Objectives:(i)To correlate the expression of IGF2BP1, IGF2BP3and ETV6 in Indian B-ALL

patients.

(ii)To study the expression of these genes in ETV6-RUNX1 positive and negative cell lines.

(iii) To study the effect of ETV6 overexpression on IGF2BP1 and IGF2BP3 levels.

Methods: RNA isolation from bone marrow samples done and 1000 ng of DNAse treated

RNA used forcDNA synthesis.ETV6-RUNX1 positive B-ALL cell line-Reh, translocation

negative cell lines-RL, and CCRF-SB, and HEK293T cell line were usedfor the study.

mailto:[email protected]

TheETV6CDS was amplified and cloned into an expression vector.The mRNA expression of

IGF2BP1, IGF2BP3 and ETV6 was studied by Real Time PCR.

Results:The expression of IGF2BP1 was significantly higher in ETV6-RUNX1 translocation

positive patients (N=16) as compared to translocation negative patients (N=24).ETV6

expression was nearly completely lost in all the translocation positive patients. The

expression of IGF2BP1 and IGF2BP3 negatively correlated with ETV6 expression.

In vitro, RL and CCRF-SB cells showed a lower expression of IGF2BP1 as compared to Reh

cells. IGF2BP3 showed a lower expression in Rehin comparison to RL and CCRF-SB cell

lines. Over expression of ETV6 in HEK293T cells resulted in down regulation of both

IGF2BP1 and IGF2BP3, whereas over expression in Reh cells resulted in down regulation of

IGF2BP3 alone.

Discussion and conclusion: A higher expression ofIGF2BP1 and low expression of ETV6 is

clearly seen in ETV6-RUNX1 translocation positive patients. Interestingly, IGF2BP3

expression appears to have a negative correlation with ETV6 expression whereas IGF2BP1

expression seems to be affected by the presence of ETV6-RUNX1 fusion gene too. ETV6 is a

well known transcriptional repressor. Data from the in-vitro and in-vivo studies points to

IGF2BP1 and IGF2BP3 being potential targets of ETV6. Further knockdown and

overexpression studies need to be done to confirm these findings.

Phd-14

Viral characteristics associated with maintenance of elite neutralizing activity in

chronically HIV-1C infected monozygotic pediatric twins

Mishra Nitesh*1, Makhdoomi Ashraf Muzamil*

1,2, Sharma Shaifali

1, Kumar Sanjeev

1, Kumar

Deepshikha1, Chawla Himanshi

1, Singh Ravinder

3, Kanga Uma

4, Das Kumar Bimal

3, Lodha Rakesh

5,

Kabra Kumar Sushil5, Luthra Kalpana

1#.

1Department of Biochemistry, All India Institute of Medical Sciences, New Delhi, India. 2Department of Biochemistry,

Government College for Women, Cluster University Srinagar, Srinagar, India. 3Department of Microbiology, All India

Institute of Medical Sciences, New Delhi, India. 4Department of Transplant Immunology and Immunogenetics, All India

Institute of Medical Sciences, New Delhi, India. 5Department of Pediatrics, All India Institute of Medical Sciences, New

Delhi, India.

Presenting Author: Nitesh Mishra, [email protected]

Corresponding Author: Kalpana Luthra, [email protected]

Abstract

Introduction

Broad and potent neutralizing antibodies (bnAbs) with multiple epitope specificities evolve in HIV-1

infected children. Chronically HIV-1 infected children that develop elite neutralizing activity are

suitable candidates to understand the mechanisms that lead to the co-evolution of virus and antibody

response targeting multiple epitopes.

Aims and objectives

Here, we evaluated the alterations in virus and antibody responses over time in chronically HIV-1C

infected monozygotic pediatric twins with potent plasma bnAbs, AIIMS_329 and AIIMS_330, who

had acquired the infection by vertical transmission. We used serum samples taken from a pair of HIV-

1C infected, genetically identical twins classified as long term non-progressors (LTNPs) - along with

the generation of HIV pseudoviruses - to study the evolutionary course of HIV in the context of an

elite neutralizing serological response.

Materials and methods

Utilizing standard HIV virus panel reflective of the global Env diversity, single base mutants, and

generating envelope pseudoviruses reflective of the circulating viral variants in the monozygotic

twins, we assessed the plasma antibody neutralizing activity, characterized the epitopes against which

the plasma antibody response was directed, identified mechanisms of viral escape from serum

neutralization as well as assessed the features associated with maintenance of elite plasma neutralizing

activity.

Results

In the current study, we studied the longitudinal development of antibody responses and its

association with the circulating viral immunotypes in a monozygotic twin pair infected with HIV

clade C viruses over a period 60 months with first sampling at 78 months post infection. Both HIV-1

chronically infected antiretroviral twins maintained CD4+ T cell levels and viral loads throughout and

their plasma antibodies showed cross neutralizing activity and targeted multiple epitopes of HIV-1

envelope glycoprotein. While the plasma neutralizing antibody(nAb) titers persisted and increased in

potency and breadth with time in AIIMS_330, the other twin AIIMS_329, showed an initial high nAb

titre that decreased in later time points Interestingly we observed higher viral diversity in the



pseudoviruses generated from AIIMS_330 with majority of the viral variants sensitive to autologous

plasma antibodies compared to AIIMS_329 in whom the circulating viral variants were resistant to

autologous plasma antibodies.

Conclusion

A longitudinal analysis of the alterations in the HIV-1 neutralizing activity of the plasma antibodies

and in the properties of the contemporaneous and evolving viruses in a pair of HIV-1 infected

genetically identical twin children showed that higher diversity of the viruses in AIIMS_330 as

compared to the AIIMS_329 viruses may have contributed to the development of potent bnAbs in

AIIMS_330 that persisted throughout. Such a pair of genetically identical twins that have presumably

been infected by the same founder virus serve as valuable biological samples to understand the

mechanisms that lead to the co-evolution of virus and host immune response.

Phd-15

EVALUATING THE ROLE OF METFORMIN ON DIFFERENTIATION AND TUMOR MICROENVIRONMENT IN HUMAN COLORECTAL CARCINOMA

Kaur, Preet Amar1; Dey, Devanjan1; Bhattacharya, Aditi1; Bansal, Aakriti2; Das, Prasenjit3; Vishnubhatla, Sreenivas4; Sen, Sudip1. 1Department of Biochemistry, AIIMS, New Delhi. 2Department of Biotechnology, Jamia Hamdard Deemed University, New Delhi. 3Department of Pathology, AIIMS, New Delhi. 4Department of Biostatistics, AIIMS, New Delhi. Presenting author: Name: Amar Preet Kaur Email: [email protected]

Corresponding author: Name: Sudip Sen Email: [email protected]

Introduction: Metformin, a well known antidiabetic drug has been repurposed as an anticancer drug. In this study, the effect of metformin on differentiation and tumor microenvironment of colorectal cancer was evaluated. Aim and objectives: Aim: To study the association between differentiation and tumor microenvironment in human colorectal carcinoma cancer cell lines and tumors. Objectives: 1. To study the effect of metformin as a differentiating agent in human colorectal carcinoma cell lines. 2. To analyze changes in tumor microenvironment interacting proteins in metformin treated cancer cell lines and human colorectal carcinoma samples of different histopathological grades.

Material and methods: Maximum tolerable non-toxic dose of metformin on HCT116 (poorly differentiated) and HT29 (moderately differentiated) colon cancer cells was determined by MTT assay. Cells were treated with the selected doses (1, 2.5 and 5 mM) of metformin for 2 weeks. Metformin’s ability to induce differentiation in cancer cells was assessed by analyzing Cytokeratin 20 (CK20) by flow cytometry and CDX1 (transcription factor for CK20), by RT-QPCR. Analysis of apoptosis was done by flow cytometry. Expression of Ki67 (proliferation marker) and Extracellular matrix (ECM) proteins (COL3A1, COL4A2, COL5A2, ITGA6, VTN, LAMC3 and TNXB) were studied by RT-QPCR. Immunohistochemistry (IHC) was performed in human colorectal cancer tissue of different histopathological grades to evaluate the expression of the ECM proteins. Results: After two weeks of metformin treatment (1, 2.5 and 5 mM), expression of CK20 (HCT116:Control(C)-13%, 2.5mM- 33% and HT29: C-17%, 1mM- 96%, 2.5mM-99%, 5mM-98%) and CDX1(HCT116: 4.8 fold with 2.5mM, 4.1 fold with 5mM


andHT29: 4.6 fold with 1mM, 4.6 fold with 2.5mM. 4.9 fold with 5mM) were found to be increased. In HCT116 cells, the percentage of early apoptotic cells with 2.5mM (15.8%)

metformin treatment was slightly increased as compared to control (11.6%) and the percentage of late apoptotic cells was increased with 5mM (18.1%) metformin as compared to control (7.1%). In HT29 cells, the percentage of early apoptotic cells with 2.5mM (8.9%) and 5mM metformin (12.4%) treatment was increased as compared to control (2.0%).

Expression of Ki67 was decreased with higher doses of metformin (HCT116: 5mM-2.5 fold and HT29: 5mM-3.3 fold). The expression of ECM proteins were significantly decreased with metformin in HCT116(COL3A1-5 fold, COL4A2-5 fold, COL5A2-10 fold, ITGA6-11 fold, VTN-10 fold, LAMC3-10 fold and TNXB-3 fold) and HT29 (COL3A1-5 fold, COL4A2-5 fold, COL5A2-5 fold, ITGA6-10 fold, VTN-10 fold, LAMC3-3 fold and TNXB- 2.5 fold) cells. IHC corroborated this finding in the ECM of human colorectal cancer tissue sections. Conclusion: Our study highlights that Metformin may be acting on colorectal cancer cells through multiple mechanisms like inducing differentiation, thereby reducing proliferation, as evidenced by alteration in differentiation markers and cell proliferation. Alteration in the expression of ECM proteins highlights the putative therapeutic role of metformin in regulating the niche supporting these cancer cells, thereby playing an important role in modulating cancer progression and metastasis.

Phd-16

The Involvement of Iron in Neurodegeneration associated with

Friedreich’s ataxia

Deepti Pathak1, Achal Kumar Srivastava

2, Sheffali Gulati

3, M.V. Padma

2, Moganty R

Rajeswari1

1 Department of Biochemistry, All India Institute of Medical Sciences, New Delhi, India

2 Department of Neurology, All India Institute of Medical Sciences, New Delhi, India

3 Department of Paediatrics, All India Institute of Medical Sciences, New Delhi, India

Abstract

Introduction: Friedreich's ataxia (FRDA), a progressive neurodegenerative disorder caused

by trinucleotide (GAA) repeat expansion in frataxin (fxn) gene which results in decreased

levels of frataxin protein. Insufficient frataxin levels leads to iron and copper deposits in the

brain and cardiac cells, thereby, and increased oxidative stress.

Aims and Objective: To assess their cell-free levels in FRDA patients, in order to check

whether the disturbed homeostasis of iron and copper is reflected in blood plasma.

Methods: A total of hundred and twenty patients, suspected of FRDA were screened for the

genetic analysis for (GAA) repeats in the fxn gene by PCR and those (25) patients confirmed

for FRDA were recruited in the study. The total Iron and total copper concentrations were

measured in blood plasma using Nitro PAPS and Dibrom PAESA method, respectively both

in patients and age, sex matched healthy controls.

Results: The iron levels mean ± SD (6.2 ± 3.8) in plasma of FRDA patients were found to be

significantly decreased as compared to healthy controls mean ± SD (15.2 ± 4.2). A similar

trend was also observed in case of plasma copper levels in FRDA patient (8.15± 4.6) as

compared to controls (17.5± 3.40)

Conclusion: Present results clearly prove abnormal distribution of extra-cellular iron in

FRDA patients, which is in accordance with the well established fact of intracellular iron

overload, which is the key feature of the pathogenesis of this disease. This can be of

importance in understanding the pathophysiology of the disease in association with frataxin /

iron. It appears that intracellular sequestration of trace metals in FRDA patients (due to low

frataxin) results in their sub-optimal levels in blood plasma (extra-cellular) an observation

that can find prognostic application in clinical trials.

Keywords - Friedreich's ataxia (FRDA); Neurodegenerative disorder; mitochondrial iron

accumulation; Plasma iron and copper; ROS

Phd-17

Title “CHARACTERISIATION OF FAT1 PROCESSING AND ITS INTERACTION

WITH β-CATENIN IN GLIOMA”

Gowda H Srinivas1, Gupta Yakhlesh

1, Malik Nargis

1, Goswami Sanjeev

1, Chattopadhyay

Parthaprasad1, Sarkar Chitra

2, Suri Ashish

3, Gupta K Deepak

3, Sinha Subrata

1, Chosdol

Kunzang1.

1Department of Biochemistry, AIIMS; New Delhi, India;

2Department of Pathology, AIIMS,

New Delhi; 3

Department of Neurosurgery, AIIMS, New Delhi;

Presenting Author:

Name : Dr Srinivas H

Email : [email protected]

Corresponding Author:

Name : Dr Kunzang Chosdol


Introduction:

FAT1 is a 506kDa transmembrane protein. It functions as an adhesion molecule and has

known interaction via its intracytoplasmic region with various protein along with beta

catenin. FAT1 having differential processivity and has different subcellular localisation

suggesting its intracellular signalling functions apart from being adhesion molecule.

Glioblastoma multiforme (GBM) patients have a very poor prognosis and average survival

rate is less than 18 months, so there is a need to identify new molecular therapeutic. Studying

the role of FAT1 and its interaction with beta catenin in glioma will increase our knowledge

and enable to develop a treatment option with this molecule.

Aims: To study the oncogenic role of FAT1 in hypoxia in glioma

Objectives: 1) Characterisation of FAT1 processing in glioma and 2) Interaction of FAT1

and β-Catenin in glioma.

Materials and methods:

Experiment were carried out in U87MG glioma cells maintained under normoxia (20%O2 and

severe hypoxia (0.2%O2). Immunocytochemistry, migration/invasion, western blotting and

real-time gene expression study was carried out after FAT1 modulation (FAT1 over

expressed using trunc FAT1 plasmid/ knockdown using siFAT1). Furin inhibitor treatment

(20μmol for 48hrs) was done to analysis FAT1 processing in glioma. CO-Ip experiment was

done to study the interaction of FAT1 and β-Catenin in different subcellular fraction.

Results:

We observed along with full-length FAT1 protein band (508 kDa), multiple smaller bands in

the range of 171 to 117 kDa in different cell lines. On Furin inhibitor treatment, there was

decrease in the intensity of cleaved protein bands (171 to 117 kDa 40% reduction) and

increase in the full length FAT1 protein (1.53 fold) under hypoxia. In addition, Furin

inhibitor treatment, decreased p53, HIF1α and p300 expression as well as 60% decrease in

the migration of glioma cells, reflecting the importance of FAT1 processing for its various

cellular functioning.

On exposure to hypoxia there was 1.98 folds increase in FAT1 protein and also increased

nuclear localisation on exposure to hypoxia on ICC experiment in U87MG cells. Nuclear

localisation of FAT1 on exposure to Hypoxia was reduced on treatment with Furin Inhibitors.

β-catenin has no nuclear localisation domain and FAT1 has predicted Nuclear localisation

signals in its intracytoplasmic domain. FAT1 mediated pull down showed interaction with β-

catenin in our CO-IP experiment in nuclear lysate. FAT1 knockdown decreases β-catenin

ICC signals in nucleus both under hypoxia and normoxia. We observed the interaction of β-

catenin and FAT1 in nuclear fraction of U87MG cells (CO-IP experiments) and FAT1

modulating β-catenin downstream genes. On FAT1 over expression under hypoxia, we

observed, increase in the levels of target proteins of β-catenin (c-JUN 1.63 fold, c-MYC 1.48

fold, VEGF 1.38 fold and Cyclin D 1.17 fold) compared to hypoxia pcDNA control cells and

decreased on FAT1 knockdown samples.

Conclusion: FAT1 protein expression is increased on exposure to severe hypoxia and

enhanced nuclear translocation compared to Normoxia in glioma cells. FAT1 undergo Furin

mediated proconvertase processing in glioma under hypoxia and FAT1 may act as oncogene

by aiding transportation of known oncogene β-catenin to nucleus under hypoxia condition.

Phd-18

Title: Transactivation of HnRNPD/Auf-1 by NF-κB(P65/RelA) in oral cancer

Authors: Kumar Vikas, Kumar Manish, Chauhan S Shyam

Affiliation: Department of Biochemistry, All India Institute of Medical Sciences (AIIMS), New Delhi

Name of Presenting Authors: Vikas Kumar

Mobile No: 9899480067

Email ID: [email protected]

Corresponding Author: Prof. Shyam S Chauhan


Abstract

Introduction: Head and neck oral squamous cell carcinoma (HNOSCC) is a heterogeneous group of

malignancies that arise in upper aero-digestive tract. Approximately 90% of HNOSCC are squamous cell in

origin. Regulation of mRNA stability is an important post transcriptional regulatory mechanism of gene

expression which has been implicated in tumorigenesis. Approximately 16% of mRNA contains AU rich regions

(ARE), in their 3’UTRs. Heterogeneous ribonucleoprotein D/Auf-1 is an RNA binding protein (AUBP), that binds

to 3’UTR ARE of many mRNA and modulate their decay rates by deploying mRNA degrading proteins.

Previously our laboratory has demonstrated over expression of hnRNPD in HNOSCC and established its

correlation with poor prognosis of the disease. However, till date no systematic study has been carried out to

elucidate the molecular mechanism of its over-expression in oral cancer. Therefore present study was

undertaken to elucidate the mechanism of transcriptional regulation of hnRNPD expression in oral cancer.

Objectives: PCR amplification and cloning of hnRNPD promoter region in pGL3-Basic vector followed by

demonstration of promoter activity in the cloned fragment and identify the functional transcription factor(s)

binding motif(s) on hnRNPD promoter by promoter deletion analysis.

Material and Methods: Upstream region of hnRNPD gene was PCR amplified and subjected to double

stranded DNA sequencing and the nucleotide sequence thus obtained was aligned with nucleotide sequence of

upstream region of hnRNPD gene. Fragment exhibiting 100% homology to the upstream region of hnRNPD

gene was cloned upstream to the promoterless luciferase reporter gene into pGL3-Basic vector. The resulting

vector was transfected in two different oral cancer cell lines SCC-4 and SCC-25 to assess promoter activity of

the cloned fragment. Promoter deletion analysis and ChIp assays were performed to identify the promoter

region and transcription factor(s) respectively involved in regulation of hnRNPD expression.

Results: Promoter reporter assay demonstrated significantly higher hnRNPD promoter activity in SCC4 cells as

compared to SCC25 cells. In silico analysis of cloned hnRNPD promoter region revealed many putative

transcription factors binding sites including four NF-κB1 and one NF-κB (RelA/P65) binding motifs. Deletion of



NF-κB (RelA/P65) binding motif located between -1150 and -1161 resulted in a significant decrease in hnRNPD

promoter activity. ChIP assay revealed binding of transcription factor NF-κB to this motif. Furthermore,

treatment of SCC4 cells with PDTC, a specific inhibitor for NF-κB specific abolished NF-kB binding to this motif

with a concomitant decrease in the level of hnRNPD in oral cancer cells.

Conclusions: Results of the present study for the first time establish the involvement of NF-κB in

transcriptional up regulation of hnRNPD

Phd-21

ROLE OF VITAMIN D AND ITS ASSOCIATED MOLECULES IN INNATE IMMUNITY

MODULATION IN PULMONARY TUBERCULOSIS

Sudhasini Panda1#, Ambrish Tiwari1, Kalpana Luthra1,SK Sharma2, Archana Singh1*

1Department of Biochemistry, All India Institute of Medical Sciences, New Delhi -110029, India.

2Department of Medicine, All India Institute of Medical Sciences, New Delhi -110029, India.

#Presenter:[email protected]

*Corresponding author:[email protected]

Introduction: Innate immunity plays an important role in pathophysiology of tuberculosis which is

influenced by various host factors. One such factor is vitamin D that along with its transport protein

vitamin D binding protein (VDBP) and its nuclear receptor vitamin D receptor (VDR), may play a

role in altering the host defense against Mtb via production of cathelicidin (antimicrobial peptide) and

regulating the expression of inducible nitric oxide synthase (iNOS) required for production of Nitric

oxide (NO) which also has bactericidal effect. With this background we attempted to investigate the

levels of Vitamin D and NO along with their associated molecules in tuberculosis patients and

household contacts as compared to healthy controls and assess the implication of these findings in

susceptibility to tuberculosis (TB).

Materials and methods: 80 active TB patients, 75 household contacts and 70 healthy controls were

taken. VDR, VDBP and iNOS mRNA levels were studied using real time PCR. Serum VDR,

cathelicidin, iNOS levels were measured using ELISA. Serum Vitamin D levels were measured in

serum samples using chemiluminiscence based immunoassay. NO was measured using colorimetry

based kit.

Results: VDR and iNOS mRNA levels were found to be lower in active TB group compared to

household contacts and healthy controls (P=0.0001 and 0.005 respectively). Though insignificant,

expression of VDBP mRNA was lower in active TB group as compared to household contact and

control group. The serum levels of Vitamin D were also found to be lower in active TB group as

compared to healthy controls (P =0.001). Levels of cathelicidin and NO was higher in patient group as

compared to other groups (p=0.01 and 0.5 respectively). However, the expression of VDR and iNOS

and levels of vitamin D was significantly (P < 0.05) higher in household contacts compared to both

active TB and healthy control groups.

Conclusion: Higher levels of Vitamin D along with VDR and iNOS expression in household contacts as

compared to patients suggest that vitamin D might have protective role against TB which prevents

activation of the disease. From our data, we can conclude that decreased vitamin D levels could be

implicated in disease progression. Supplementation of vitamin D and arginine alone in household

contacts of TB and as an adjuvant therapy along with anti tubercular treatment in active TB should

be evaluated to assess their therapeutic potential.

Phd-22

Targeting of versican by microRNAs inhibit multiple myeloma

progression: A novel potential therapeutic strategy

1Gupta Nidhi,

1Kumar Raman,

2Seth Tulika,

3Garg Bhavuk,

1*Sharma Alpana

1Department of Biochemistry, All India Institute of Medical Sciences, New Delhi, India

2Department of Hematology, All India Institute of Medical Sciences, New Delhi, India

3Department of Orthopedics, All India Institute of Medical Sciences, New Delhi, India

Presenting Author: Nidhi Gupta

[email protected]

*Corresponding Author: Prof. Alpana Sharma

Email id: [email protected]

ABSTRACT

Introduction: Multiple Myeloma (MM) is second most common hematological malignancy

characterized by uncontrolled proliferation of abnormal plasma cells in bone marrow (BM).

These myeloma cells require BM niche consisting of proteoglycans, cytokines and growth

factors, etc. for their growth. One of the chondroitin sulfate proteoglycan, Versican (VCAN)

has gained consideration in milieu of solid tumors where it has shown to promote tumor

progression but there is dearth of literature in hematological malignancies including MM.

Aim & Objectives: Hence, we aim to study the involvement of VCAN and its regulation by

microRNAs in MM. To achieve this hypothesis, expression of VCAN and microRNAs (miR-

144, miR-199 and miR-203) have been studied in MM patients and cell lines. Further,

involvement of VCAN in myeloma pathogenesis has been assessed by certain in vitro

experiments followed by elucidating the regulation of VCAN by microRNAs.

Materials and Methods: 30 MM patients & 20 controls were recruited and BM Stromal

Cells (BMSCs) were harvested by primary culture. Molecular expression of VCAN and

microRNAs (miR-144, miR-199 & miR-203) were examined in study subjects and MM cell

lines (RPMI8226 & U266). VCAN levels were correlated with microRNAs by Spearman

correlation analysis. Conditioned medium (CM) of BMSCs was examined for presence of

VCAN by ELISA and its effect on myeloma pathogenesis was studied in presence or absence

of VCAN antibody. Further, signaling pathways altered by VCAN were identified. VCAN

regulation by microRNAs was then assessed by transfecting miR mimics (miR-144 & miR-

199) in primary BMSCs and CM thus obtained was supplemented to MM cells to investigate

alteration in effects caused by knockdown of VCAN by miR mimics. Significance was

determined using Mann-Whitney U test and student’s t-test.

Results: Relative mRNA expression of VCAN was found significantly higher in MM

patients especially in bone marrow stroma while relative microRNA expression was

significantly lower in patients. VCAN levels showed negative correlation with microRNAs.

VCAN being produced in stroma found at lower levels in MM cell lines. Moreover, BMSCs-

CM showed presence of VCAN which upon supplementing to MM cells alter parameters in

favour of myeloma progression, however, this effect was neutralized by VCAN antibody.

The downstream signaling of VCAN was found to activate FAK and STAT3 which subsides

by VCAN antibody. The transfection of miR-144 and miR-199 mimics in BMSCs leads to

significant reduction in levels of VCAN. The effect caused by VCAN on myeloma cells has

also been neutralized by miR mimics.

Conclusion: Augmented levels of VCAN in BM of patients imply its involvement in BM

niche of MM. The neutralization of oncogenic effect of BMSCs-CM by VCAN antibody

affirms plausible role of VCAN in progression of MM. Moreover, myeloma promoting effect

of VCAN has been reversed by microRNA (miR-144 and miR-199) mimics. These findings

open up new avenues for exploring VCAN as a novel therapeutic target and microRNAs as a

mean to target VCAN for treatment of MM in clinical settings in future.

Phd-28

FAT1 gene regulates Programmed cell death 10 (PDCD10) via miR-221 in glioblastoma

Nargis Malik, Parthaprasad Chattopadhyay, Chitra Sarkar, Ashish Suri, Subrata Sinha, Kunzang Chosdol Background:-FAT1 gene is localized on chromosome 4q35.2 coding a 506KDa transmembrane

protein and functions as an oncogene or tumor suppressor depending on tissue types in human

cancers. Our lab has identified that FAT1 has oncogenic role in glioblastoma (GBM). PDCD10 gene

encodes an evolutionarily conserved protein that is widely expressed in almost all human tissues. Here

in our study we are characterizing the role of FAT1 gene in primary brain tumors and in the regulation

of miRNAs and its target gene inGBM.

Methodology:-In order to define molecular crosstalk among factors driving glioma progression, we

used knockdown method in glioma cell lines followed by expression analysis by Q-PCR. FAT1

knockdown was done usingFAT1 specific siRNA and mRNA and miRNA expression

analysis by specific primers in GBM cell lines. Expression and Spearman correlation analysis

of FAT1 with miR-221-3p and PDCD10 was done in GBM tumor samples (n=30). In a

sequential bioinformatics study, we analyzed TCGA GBM dataset for FAT1 and PDCD10

expression and perform spearman correlation in GBM tumors (n=430) as compared to normal

brain (n=10).

Results:-On FAT1 knockdown, using FAT1 specific siRNA we observed significantly

decreased expression of miR-221/222-3p. In-silico analysis recognized, PDCD10as a

potential targets of miR-221/222-3p.Furthermore, FAT1 knocked-down cells showed

significantly increased expression of PDCD10 in all studied glioma cell lines. In order to

validate our in-vitro observation and its clinical relevance, we have done expression and

correlation study in GBM tumor samples. Weobserved significantpositive spearman

correlation between FAT1 and miR-221-3p (r=0.5669, p≤0.0011)and negative correlation of

FAT1 with PDCD10 (r= -0.3492, p≤0.0585),) and miR-221-3p with PDCD10 (r=0.526,

p≤0.0028). In TCGA database we have observed negative correlation between FAT1 and

PDCD10(r=-0.4209, p≤0.0001).These results propose that FAT1 regulating the expression of

miR-221-3p leading to downregulation of PDCD10 in GBM cell lines and GBM tumors.

Conclusion:-Our in-vitro and GBM tumor data for the first time suggesting FAT1 to be a

novel molecule regulating the expression of PDCD10 via miR221-3p in GBM

PDCD10 gene encodes an evolutionarily conserved protein that is extensively expressed in nearly all

human tissues

Phd-82

Title - Silver nanoparticle enhance tumor associated macrophage function

and decreases tumor cell proliferation

Name of the authors - Gautam Kumar Pramod, Singh Khushwant, Anita, Garg Drishti, Bukhari

Jehangir.

Affiliation:

Presenting Author: Name - Khushwant Singh

Email – [email protected]


Name – Dr. Pramod Kumar Gautam

Email - [email protected]

Abstract Introduction: Tumor-associated macrophages (TAMs) or M2 macrophages (alternatively activated) that

constitute about 10–20 % of total tumor cell mass and these cells serve for the tumor as slaves.

Tumors burden are generally characterized by nutrient deprivation, hypoxia. Tumor

microenvironment is housed with both malignant and non-malignant cell types. Tumor cells

release several immunosuppressive factors, pro-angiogenic cytokines and growth factors that

stimulate endothelial cell proliferation and promote the formation of the differentiated capillary

tube. These tumor-promoting factors further down-regulate macrophages function and change

their functional phenotype M1 to altered M2 phenotype.

Aims & Objectives: Silver has been a widely used molecule in therapeutics due to its potent anti-bacterial, anti-fungal

and anti-viral activity along with its ability to have immunomodulatory effects in the form of

silver nanoparticles. It is speculated that the potential of silver could be due to the nanoscale size

and shape of AgNPs

Materials and methods: AgNPs were synthesized using Silver nitrate by added drop wise to the solution of stirred and

cool sodium citrate of double distilled water and UV spectra were taken at a different time

interval. Further FT-IR, XRD, TEM was performed to determine the functional group upon

synthesis. Different assay like MTT assay, Nuclear morphology assay, microscopy, DNA

fragmentation assay, tumor associated macrophage Isolation, purification etc. were performed.

Results: The results showed that synthesized silver nanoparticles have anticancer activity on Daltons

Lymphoma (DL) cell line with the IC50 value of <56μg/ml confirmed by MTT assay. DL cells

treated with the AgNPs showed reduced cell viability, altered nuclear morphology, typical

apoptotic DNA ladder, and reduced mitochondrial membrane potential. It was also found that



AgNPs treatment enhanced the M2 phenotype function of macrophage harvested from tumor-host

and induced the ROS expression, fusion, adhesion, and phagocytic function.

Conclusion:

From the above finding, it can be concluded that AgNPs have the potential to decrease the

proliferation of tumor cells and enhanced the macrophage potential.

Phd-84

Title: HDL abnormalities in cardiovascular disease: Quality or Quantity?

Name of the authors: Thakkar Himani1, Vincent Vinnyfred

1, Roy Ambuj

2, Singh Sandeep

2,

Ramakrishnan Lakshmy3, Singh Archna

1

Affiliation:

1Department of Biochemistry, AIIMS, New Delhi

2Department of Cardio-Thoracic Science Centre, AIIMS, New Delhi

3Department of Cardiac Biochemistry, AIIMS, New Delhi

Presenting Author:

Name: Himani Thakkar



Name: Dr. Archna Singh


Introduction:

Failure of several HDL modulating drugs in reducing the cardiac risk has raiseddoubts on

thewell accepted paradigm of HDL cholesterol’s roleas a negative risk factor for

CardiovascularDisease. Since increasing HDL-C levels alone does not appear to enhance

thebiological activities of HDL, a betterunderstanding of HDL structure-function

relationships and assessment of HDL functionality is suggested fordesigning interventions

that can produce benefit. Since, Indians tend to develop CVD at a younger ageand appear

predisposed to risk factors (low HDL-C levels) associated with acute coronary syndromes,

we assessed HDL related functional parameters in this group.

AIM:

The aim of the study was to evaluate HDL functionality in patients with ACS and its

modulation after standard therapy including statins.

OBJECTIVES:



To analyse the cholesterol efflux capacity (CEC) and antioxidative activity as reflected in

Paraoxonase 1(PON1) activity of HDL in patients of acute coronary syndrome at baseline

and after six months of therapy including statins.

MATERIALS AND METHODS:

Statin naïve ACS (n=151) and control subjects (n=110) were enrolled in the study.

Macrophage-specific efflux capacity of HDL was measured using fluorescently labelled

(BODIPY) cholesterol. Uptake of fluorescently labelled cholesterol by HDL from THP-1

derived macrophage foam cells was analysed by measuring the change in fluorescence

intensity (FI) of medium with and without HDL. The difference in FI of the medium

containing HDL (index of cholesterol efflux capacity) from subjects and controls was

compared. Paraoxonase and arylesterase activities of PON1 enzyme were assessed by

spectrophotometric methods. Cholesterol efflux capacity and PON1activity was investigated

after 6 months of statin therapy in ACS patients (N=100).

RESULTS:

Of the total 151 ACS patients, 132 (88%) were men whereas in control group, 65 (59%)

of110 subjects were males. Mean age of the subjects in control group was 43.6 ± 10.5 years

and in ACS group was 51.7 ± 11.4 years. Patients presenting with ACS had significantly

lower HDL-C (45.4 ± 9.6 vs 40.2 ± 9.6, p <0.0001) and apolipoprotein A-I (142.1 ± 43.3 vs

112.5 ±27.3, p <0.0001) levels as compared to control subjects. ACS patients had

significantly higher apo B levels as compared to controls; however, noo significant

differenceswere observed in total cholesterol (TC), LDL-C, very low density lipoprotein

cholesterol (VLDL-C) and triglycerides (TG) between the two groups.

In the study, ACS patients had significantly lower cholesterol efflux capacity (CEC) and

antioxidative capacity of HDL compared to controls, which improved significantly after six

months of treatment. Higher levels of CEC, Paraoxonase 1 activity and apolipoprotein A-I

were associated with lower odds of development of ACS. The association was significant

even after adjusting for cardiovascular risk factors and HDL-C levels. CEC was positively

with apolipoprotein A-I levels in ACS subjects at baseline and after treatment

CONCLUSION:

The significant association of lower Cholesterol efflux capacity and antioxidative activity of

HDL with increased risk of development of ACS and improvement in HDL functionality

after pharmacological intervention indicates that HDL functionality would be a better

measure of cardiovascular risk than measuring static mass of HDL-cholesterol.

Phd-89

Unraveling the role of Gamma Delta T cell subsets: Key players in the

immunopathogenesis of Pemphigus Vulgaris

DasDayasagar,KV Santosh, AravaSudheer, KhandpurSujay, Sharma Alpana

Presenting author:

Dayasagar Das- PhD Student


Corresponding author:

Prof. Alpana Sharma


Introduction:

Autoimmune diseases are one of the challenging puzzles in immunology due to its

multiple etiologies, complex mechanisms and associated severity. Breach in tolerance

and anomalous T cell function are important phenomenon associated with autoimmune

diseases. Pemphigus Vulgaris (PV) is a severe form of autoimmune blistering disease

involving skin and mucosa. Pathophysiology of PV is due to the formation of auto-

antibodies against desmoglein3 (Dsg3) and anomalous T cell function.Gamma Delta T

cells (γδ T cells) are unique multifaceted T cells that maintain the immune surveillance at

the epithelial surface.They have unique property to recognize the antigens in MHC

independent manner and activating the adaptive arm of immune system. These cells have

found to be involved in several autoimmune diseases. However, their role and plasticity

are not yet characterized in the pathogenesis of PV.

Aims & Objectives: To explore the role of γδ T cell subsets and their plasticity in the

immunopathogenesis of PV.

Materials and methods: 30 active PV patients confirmed by clinically (H&E, DIF

staining) and 30 controls were included.Frequency of γδ-T cell subsets were assessed by

Flowcytometry. Patient and control derived γδ-T cell were isolated by MACS and

primary culture was done. Circulatory and culture levels of γδ-T cell associated markers

were estimated by ELISA. Polarization potential and the functional markersrelative m-

RNA was done by cytotoxicity assay &real time PCR respectively. Tissue localization of

γδ-T cell subsets was confirmed by Immunohistochemistry.

Results:Primary culture of γδ T cells isolated from the PV patients showed diverse

phenotypes (γδ1, γδ2, γδ17, γδreg) and functionality, such as migration, multiple cytokine

productions. We have observed the significantly higher frequency of pan γδ T cells (6.7%

vs. 4.2%) in PV patients. We also found an increased frequency of IL-17(16% vs.4.6%)

and IFN-γ (36% vs.21%) producing γδ T cells in the circulation of patients. Cytotoxic

activity of γδ T cells from PV patients (26±11%) was observed to be higher as compared

to control (12±7%) and the Th1 polarization (IFN-γ) suggesting the potential role of these



cells in the pathogenesis of PV. The molecular expression (mRNA) of γδ T cell subsets

related markers (IFN-γ, IL-1, IL-17, RORγt, CD27, and CD70) were significantly

elevated in patients. Further, tissue localization of γδ T cells and its associated receptor-

ligands were found to be overexpressed in the skin of PV patients.

Conclusion: Exploring γδ T cell subsets in PV has revealed functional facets of T cells

and their crucial involvement. Further exploitation of these potential markers associated

with γδ T cell can be utilized to develop newer diagnostic tools and novel therapeutics for

controlling autoimmune skin diseases.

Phd-98

Title: Elevated expression of Dipeptidyl Peptidase III correlates with the Tumor Infiltrating

Lymphocyte(TILs) composition in multiple cancers.

Authors: Arora Mohit1,Gupta Jasmeen

1, Prajapati Chand Subhash

2,Chauhan Singh Shyam

1*.

1Department of Biochemistry, All India Institute of Medical Sciences (AIIMS), New Delhi.

2Department of Biochemistry and Molecular Genetics, University of Virginia, USA.

Presenting Author:

Name: Mohit Arora



Name: Dr.Shyam Singh Chauhan


Abstract

Introduction: Immune escape is a hallmark of cancer. Recent evidence suggests that Tumor

Infiltrating Immune Cells (TIICs) composition might play critical roles in the pathogenesis

and treatment of several cancers. Critical determinants of tumor immunogenicity include

expression levels of antigenic peptides in tumor cells, engagement of immune cells into the

tumor tissue, presentation of antigenic peptides to immune cells and expression of immune

checkpoint proteins. Concerning this, our lab previously suggested a potential role of

Dipeptidyl Peptidase III (DPP3), a ubiquitously expressed N-terminal exopeptidase, in

degrading antigenic peptides and thereby suppressing immunity against tumor antigens. Later

on, other research groups reported that DPP3along with another peptidase THOP1 potentially

cleave antigenic peptides and mediates inhibition of T cell cross-priming, thereby supporting

our hypothesis. While elevated DPP3 expression in the tumor as well in serum has been

reported in some cancers, its clinical significancehas not been evaluated. Also, the relation

between DPP3 expression and immune cell composition in cancer has not been studied.

Aim and Objectives: To study the expression of DPP3 in different cancers and its relation

with tumorinfiltrating immune cell composition.

Objective 1: To measure DPP3 mRNA level in different cancer types.

Objective 2: To assess the potential of DPP3 in degrading antigenic peptidesin vitro.

Objective 3: To correlate DPP3 expression and tumor immune cell infiltration.

Materials and Methods:



To assess the relative mRNA levels between normal tissues and tumor tissues, we performed

gene expression analysis using RNAsequencing datasets of patients of several cancer types

from The Cancer genomic Atlas (TCGA) using standard workflow of data analysis tool,Gene

expression Profiling Interactive Analysis(GEPIA). We further assessed the potential of DPP-

3 in degrading antigenic OVA and NP peptides in vitro. Toexplore the clinical significance of

DPP-3 expression in modulating antitumor immunity, we used Tumor IMmune Estimation

Resource(TIMER), which is a tool based on deconvolution algorithm using RNA-sequencing

data to extract information about the relative abundance of different immune cells within the

bulk tumor samples of TCGA. Correlation between DPP3 expression and panel of immune

cell signature genes in some cancer types were also quantified using TIMER.

Results:DPP3 was found to be differentially expressed in multiple cancers. Its high

expression in colon and stomach cancer correlated with favorable overall survival.DPP-3 was

found to degrade antigenic peptides, which was inhibited after addition of the aminopeptidase

inhibitor. Further, DPP3 expression exhibit a strong negative correlation with the number of

TIICs including B cells, CD8+ T cells, CD4+ T cells, macrophages, neutrophils, and

dendritic cells in multiple cancers.

Conclusion:

Current data supports DPP-3 as a potential biomarker for immune infiltration in cancer and as

a target for enhancing antitumor immunity.

Phd-103

Acute Febrile Illness associated malaria co-infections in tropical

diseases: A spectrum and incidence in Indian population

Atreyi Pramanik1*, Charandeep Kaur1*, Rajendra Mandage1*, Vinod Kumar2, Shounak Saha3, Adarsh

Singh3, Manish Soneja2, Pragyan Acharya1#,

*These authors have contributed equally

# Corresponding Author: [email protected] (Lab 3002, Department of Biochemistry,

Teaching Block, AIIMS, Ansari Nagar, New Delhi – 110029)

Author Affiliations: 1 Department of Biochemistry, All India Institute of Medical Sciences, New Delhi, India

2 Department of Medicine, All India Institute of Medical Sciences, New Delhi, India

3Department of Biotechnology, Indian Institute of Technology, Kharagpur, India

Background Acute Febrile Illnesses (AFI) is one of the causes of morbidity and mortality in Indian patients and

might be allied with other co-infections in tropical diseases. However, there is little information

available on co-infections incidence. Therefore, in present study, we aim to investigate spectrum and

incidence of AFI associated mono and co-infections in tropical diseases such as malaria, dengue,

scrub typhus, leptospirosis and chikungunya in Indian populations.

Method Patients having fever for more than a week were recruited as AFI cases in our study. A detailed PCR

characterization was performed on 99 AFI samples for screening of its causative agents, such as

Plasmodium falciparum (Pf), Plasmodium vivax (Pv), Plasmodium malariae (Pm), Plasmodium ovale

(Po), and Plasmodium knowlesi(Pk) along with common co-endemic and co-seasonal pathogens like

Dengue virus (DENV), Chikunguniya virus (CHIKV) Orientia tsutsugamushi (causative agent of

Scrub typhus) and Leptospira (causative agent of leptospirosis).

Results Dengue-malaria co-infection was found to be the major group in our cohort (44%, 29 out of 66) as

compared to malaria and its intra-species infections (33%, 22 out of 66). Further analysis of Dengue-

malaria co-infection revealed that Dengue subtype 4 (DENV4) has a high alliance with mild malaria

(MM) (17 out of 33, 51%), whereas P. vivax severe malaria was strongly associated with P. knowlesi,

Leptospira and Dengue virus co-infections.

Conclusions Based on the above results, it is recommended that a complete screening of multiple pathogens panel

should be conducted on AFI patients for proper and accurate diagnosis. In particular, Plasmodium

species especially P. knowlesi, Dengue virus, O. tsutsugamushi and Leptospira should be performed

along with a primary diagnosis such as malaria. Such retrospective studies are expected to aid in

understanding the range of co-infections in Indian population where malaria and dengue are co-

endemic.

Keywords: AFI, Malaria, Dengue, Co-infections, Mixed-infection

Phd-104

Title: FAT1, a novel regulator of YAP1- effector molecule of Salvador-

Warts-Hippo (SWH) pathway, in human Glioblastoma

Gupta Yakhlesh1, Shivajirao Sachin Shelke

1, Irshad Khushboo

1, Dikshit Bhawana

1,

Srivastav Tapasya2, Chattopadhyay Parthaprasad

1, Sinha Subrata

1 & Chosdol Kunzang

1

1- Department of Biochemistry, All India Institute of Medical Sciences, New Delhi-110029, India.

2- Department of Genetics, University of Delhi South Campus, New Delhi-110021, India.

Presenting Author:

Name: Yakhlesh Gupta



Name: Kunzang Chosdol


Introduction: Glioblastoma(GBM) is lethal brain tumor arising from supporting cells of

brain. We has recognized the oncogenic role of FAT1 gene in GBM, regulating inflammatory

and hypoxic microenvironment of the tumor as well as migratory/invasive properties of

tumor cells. In Drosophila, fat, the ortholog of FAT1, is known to regulate the Salvador-

Warts-Hippo (SWH) pathway, but its role in human is not clear. Here, we have analyzed the

effect of FAT1 on SWH pathway in glioma.

Aims and Objectives:

AIM: Effect of FAT1 as a upstream regulator of SWH pathway in human glioblastoma.

Objectives:

1. To study the effect of FAT1 knockdown on cell morphology & survival of glioma cells

2. To study the effect of FAT1 knockdown on expression of SWH pathway molecules

3. To study the effect of FAT1 knockdown on interaction of YAP1 with TEAD1

4. To study the Effect of FAT1 knockdown on subcellular localization of YAP1 and p-YAP1.

Materials and Methods: Glioma cell lines (U87MG, U373, A172, GOS3 and SW1088)

were transfected with FAT1 specific siRNA/control siRNA and analyzed the expression of

SWH pathway molecules by qPCR/Western blot. Protein-protein interactions were analyzed

by Co-immunoprecipitation (Co-IP) after over-expression of YAP1 (wild-type and mutated)

and TEAD1 with and without FAT1 knockdown. Sub-cellular localization of proteins was

analyzed by Confocal microscopy.

Results: The mRNA expression of FAT1 and SWH pathway molecules (MST1, LATS1,

LATS2, YAP1 and TEAD1) was highest in U87MG cells followed by A172, U373MG and

GOS3. After FAT1 knockdown, the mRNA expression of MST1 and BIRC2 were

significantly decreased with no change in the levels of LATS1, LATS2, YAP1, TEAD1 and

BIRC5. At protein level, increased YAP1 and phospho-YAP1 was observed after FAT1

knockdown with increased total as well as phospho-YAP1 in the cytoplasmic extract as

compared to the nuclear extract. There was significant reduction in the interaction between

YAP1 and TEAD1 in siFAT1 treated cells as compared to siControl treated cells.

Conclusion: On FAT1 knockdown, we found (i) increased YAP1 protein level, could be by

increasing the protein stability as no change was observed at the mRNA level (ii) increased

the phospho-YAP1 level as it relieves the inhibitory effect on YAP1 phosphorylation (iii) it

affects the sub-cellular localization of YAP1 by retaining YAP1 in the cytosol and thereby

(iv) decrease in the YAP1:TEAD1 interaction with decreased expression of their target gene

Birc2.

This finding of the effect of FAT1 on YAP1 in GBM is novel with features pointing towards

the oncogenic role of FAT1 by regulating YAP1 sub-cellular localization and co-

transcriptional activity independent of SWH pathway.

Phd-109

Role of IGF2BP3 in migration and invasion of epithelial cancer

cell lines

Sanjeev Goswami, Gunjan Sharma, Elza Boby, Parthaprasad

Chattopadhyay, Jayanth Kumar Palanichamy

Department of Biochemistry, All India Institute of Medical Sciences, New Delhi, India

INTRODUCTION

RNA Binding Proteins (RBPs) play crucial role in biogenesis, stability and functioning of

RNAs. Insulin like

growth factor binding protein 3(IGF2BP3) is an oncogene correlated with poor prognosis and

metastasis in

cancers but the mechanism of oncogenesis and metastasis is not known. We try to address

these lacunae.

AIM AND OBJECTIVES

To study the role of IGF2BP3 in migration and invasion of epithelial cancer cell lines by

correlating the

expression of IGF2BP3 and its target with the migration and invasion.

MATERIALS AND METHODS

RNA isolation and cDNA synthesis followed by real time PCR for IGF2BP3 ant its target

expression in

epithelial cell lines done. Migration and invasion assays were also done. The experiments

were repeated after

IGF2BP3 knockdown by specific siRNAs.

RESULTS

IGF2BP3 and its targets (CD44, MALAT1) are highly expressed in HeLa and SiHa in

comparison to MCF7.

Knockdown of IGF2BP3 leads to a decrease in the expression level of CD44, MALAT1 and

decrease in the

migratory and invasive potential of HeLa and SiHa.

CONCLUSION

High IGF2BP3 expression correlates with high migration and invasion potential of HeLa and

SiHa compared to

MCF7. Decrease in expression level of targets after IGF2BP3 knockdown implies their

stabilisation by

IGF2BP3. Decrease in migration and invasion of HeLa cells after IGF2BP3 knockdown

implies its role in

stabilizing pro-migratory transcripts.

KEY WORDS

RNA binding protein, IGF2BP3, CD44, MALAT1.

Phd-110

Title: “Feasibility of measuring sodium, potassium and creatinine from urine sample on dried

filter paper”

Author details:

Mohamad Tarik1, Lakshmy Ramakrishnan1, Ritvik Amarchand2, Harshal R. Salve2, Prashant

Mathur3, Pradeep Joshi4, Anand Krishnan2

1. Department of Cardiac Biochemistry, All India Institute of Medical Sciences, New Delhi,

India.

2. Centre for Community Medicine, All India Institute of Medical Sciences, New Delhi,

India.

3. National Centre for Diseases Informatics & Research, Indian Council of Medical

Research, Bangalore, India.

4. World Health Organization, New Delhi India.

Presenting Author

Name: Mohamad Tarik


Corresponding Author

Name: Lakshmy Ramakrishnan


Abstract:

Introduction: High prevalence of hypertension has been attributed to high dietary salt intake,

which can be monitored by measurement of urinary sodium in 24hours/spot urine in a

laboratory.

Dried urine on filter strips, if found suitable, would provide a convenient alternative that

would

circumvent the need to transport urine samples in a cold chain.

Aim & Objective: The objective of this study was to standardize estimation of sodium,

potassium and creatinine in dried urine strips (DUS) and to validate the measurement by

comparing with levels of analytes in liquid urine.

Materials & Methods: Urine was collected in filter paper strips, dried at room temperature

and,

eluted prior to estimation of sodium, potassium by ISE method and creatinine by jaffe’s

method.

After standardization of measurement in DUS, the method was validated by comparing

values

obtained in 138 urine samples by DUS method with that obtained in liquid urine sample.

Correlation coefficients were computed and bland Altman was plotted to assess agreement

between the two methods. . We also assessed storage stability of these analytes after one year

of

storage of dried urine on filter paper at 4⁰C.

Results: The mean recovery of sodium, potassium and creatinine was >95% from DUS

samples.


The Limit of detection (LOD) by DUS for sodium was 0.95 mmol/L, potassium 0.47mmol/L

and

creatinine 0.59 mg/dl. Intra-assay CV was < 5% and the inter-assay CV was <7.0% for all

three

analytes. Correlation (ICC) between DUS assay and liquid urine was 0.934 (95%CI 0.909-

0.953;

p<0.0001), 0.948 (95%CI: 0.928-0.963; p<0.0001) and 0.953 (95% CI: 0.935-0.967;

p<0.0001)

respectively for sodium, potassium and creatinine. Bland-Altman analysis shows good

agreement

between the dried urine and liquid urine sample. Sodium, potassium and creatinine were

stable in

DUS during one year of storage.

Conclusion: We conclude that the sodium, potassium and creatinine are stable in dried urine

and

are readily transferable to a liquid phase for analysis offering a convenient alternative for

monitoring dietary salt intake.

Phd-125

Laboratory of Chromatin & Cancer Epigenetics

Department of Biochemistry

BESFA: Bioinformatics based Evolutionary, Structural & Functional Analysis

In-silico analysis of POTE Paralogs

Qazi Sahar1, Sharma Neetu, Karpf R Adam2, Sharma Ashok1

1Department of Biochemistry, All India Institute of Medical Sciences, New Delhi, 2FredUniversity of

Nebraska Medical Center, Omaha, NE, USA

Presenting Author: Sahar Qazi

Department of Biochemistry, All India Institute of Medical Sciences, New Delhi

[email protected]

Corresponding Author: Ashok Sharma


[email protected]

Introduction: POTE (Prostate, Placenta, Ovary, Testis, and Embryo) gene family recently

discovered paralogs belong to cancer-testis antigen family. POTE gene family is composed of

14 closely related paralogs dispersed on autosomal pericentromeric region of eight different

chromosomes in primate.

Aims & Objectives: Our basic desideratum has been epitomized below:

1. To deduce the evolutionary spectra of POTE protein paralogs.

2. Development of better and optimized tertiary structures of POTE paralogs using

homology modelling & protein threading approaches.

3. Function prediction of each POTE protein paralog.

Materials & Methodology: POTE paralogs protein sequences were first aligned by using

CLUSTAL W with default settings and then were subjected to phylogenetic analysis in MEGA

5.2 package. For structural predictions, we employed Swiss Model and MODELLER and

Phyre2 and generated models for POTE paralogs. For the best model, a critical evaluation of

the all the models was executed by -Pro Q, RAMPAGE and PROSA. ProFunc server was used

to determine the possible biochemical function of a protein from its tertiary structure.

Results: Evolutionary & structural analysis discern the functional divergence of POTE

paralogs suggesting the fact that POTE paralogs are expressed in primates, but their

orthologues are also present in several other species. The mean distance between all the

POTE proteins is 0.66 which refers to their slow evolutionary divergence. The Tajima’s χ2

test statistic was 10.29 (P = 0.00134). Further, our results point to the different protein-

protein interaction patterns between POTE paralogs and known POTE protein clients



supporting their functional divergence after POTE gene duplication. We demonstrate that

POTE protein copies in primates have diverged functionally after the gene duplication

events.

Conclusions: POTE proteins are orthologous to many different species such as: D.

melanogaster, C. elegans, fission yeast, D. discoidum, yeast and zebrafish hinting that the

POTE sequences might have undergone an adaptive divergence in the due course of

evolution. Most of the residues in POTE protein sequences are conserved, referring to low

rate of alterations/mutations. Phylogenetic analysis of the POTE paralogs indicates that

POTE E, F, I, J A and KP are clubbed together hinting at their common divergence from other

species during evolution, while POTE B1,B2,B3,C, D, G,H, M come from only primates.

Common amino acids present in all the POTE paralogs are mainly: Serine, Arginine, Lysine

and Valine respectively which may vary in their composition. Function prediction suggests

that all the POTE paralogs are mainly involved in binding like: protein, nucleotide and ATP

binding. Moreover, they are also engaged in enzyme regulation, catalytic activity,

transporter and transferase activity respectively.

Presenting Author:

Name: Sahar Qazi


E mail : [email protected]


Name: Dr. Ashok Sharma





Phd-126

Laboratory of Chromatin & Cancer Epigenetics Department of Biochemistry

All India Institute of Medical Sciences (AIIMS), New Delhi, India

Epigenetic Regulation of Pericentromerically localized Cancer-

Testis/Germline Antigen POTE and its use as diagnostic

marker/immunotherapeutic agent in Ovarian Cancer

Manav Nisha 1*

, Kundu Subhadip 1*

, Jha Sangit 1

, Quazi Sahar 1

, Sharma Neetu

1, Kumar Pawan

2, Vanamail P

3, Mathur Sandeep

4, Sharma J.B

3,

Chauhan S.S1, Kumar Lalit

2, Kumar Sunesh

3, Karpf R Adam

5, Sharma

Ashok1*

Department of Biochemistry1, Oncology, DR. BRA-IRCH

2, Department of Obstetrics & Gynaecology

3,

Department of Pathology4, Medical AIIMS, New Delhi, Nebraska Medical Center, Omaha, USA

5

Introduction: As per recent report of National Cancer Registry program of ICMR, ovarian

cancer is third leading cause of cancer related death amongst females in India. Ovarian

cancer (OC) is a “silent killer” and is comprised of a variety of different subtypes i.e. High-

grade serous (HGS- OvCa), endometrioid, mucinous, and clear cell tumors. We are primarily

focused on cancer epigenetics and deciphering the epigenomic mechanisms with an aim to

develop new epigenetic therapeutic drugs. The recent discovery of Cancer Testis/Germline

(CT/CG) antigen expression in cancer suggests a strong link between gametogenesis and

carcinogenesis. The research idea is to investigate HGS-OvCa in pursuit of developing a

new and highly needed biomarker and immunotherapy for OC. We are indulging to open the

new avenues for CT/CG antigens i.e. POTE antigens for cancer immunotherapy against

gynaecological cancers. POTE (Prostate, Placenta, Ovary, and Testis-expressed) is a

recently discovered gene family consisting of 14 autosomal and pericentromerically localized

cancer-testis/germline antigen genes. Epigenetic modulatory agents robustly promote

expression of CG antigens, as well as the class-1 histocompatibility complex (MHCI). Thus,

we are exploring the possible clinical use of epigenetic modulators to augment the

immunotherapeutic potential of POTE family antigens, and examining how this will ultimately

improve strategies for cancer detection and treatment.

Our overall objectives are:

1. Analyzing the POTE as a New Biomarker for Ovarian Cancer: Harnessing its Clinical

Significance for Personalized Therapy

2. Development of Novel Immunotherapy based Strategies Targeting New Cancer-

Germline Antigen POTE-Paralogs in Human Epithelial Ovarian Cancer

3. To study the Epigenetic mechanism of regulation in Pericentromeric localized

Cancer-Testis/Germline Antigen POTE Expression in Ovarian Cancer

Methodology: Affymetrix HG 1.0ST microarrays and RT-qPCR were used to determine

POTE gene expression in normal ovary (NO) and OC tumors. RNA-seq, methyl-Seq and

Pyrosequencing was used for DNA methylation and POTE gene expression analysis

fallopian tube epithelia (FTE) and OC.

Results: Recently, we defined a DNA hypomethylation phenotype in EOC that consists of

hypomethylation of global genomic DNA and a family of genes known as cancer germline (CG)

antigens. POTE gene expression levels were highly elevated in OC tumors showing global

hypomethylation of LINE1 elements, as compared to EOC tumors with hypermethylation of LINE1

elements, linking global DNA methylation status to POTE expression. POTE gene knockdown in OC

cell lines resulted in significantly reduced cell migration and cell invasion. POTE genes are widely

expressed in OC, and are significantly increased in tumors displaying global DNA hypomethylation.

We hereby explore the role of 3D nuclear architecture, lamina associate domain (LADs) interactions

to understand the molecular mechanism of activation of pericentromeric localized genes.

Conclusion: As POTEs are specifically localized on the pericentromeric regions, we will be able to

explain the regulation of the pericentromeric localized genes at a known repressed locus, and their

escape from the repression during cancer. We believe that outcomes of this project will increase our

understanding about molecular clinicopathological mechanism of ovarian cancer that will also help to

develop better cancer-testis/germline POTE based therapeutic regimen for relapsed patient in future.

Presenting Author:

Name: Ms. Nisha Manav


E mail : [email protected]


Name: Dr. Ashok Sharma





Phd-134

Identifying molecules to distinguish between colorectal cancer stem cells and normal

colonic stem cells to develop novel diagnostic and prognostic markers

Bhattacharya Aditi1, Manhas Janvie1, Dey Devanjan1, Bhat Muzaffar2, Das Prasenjit3, Pal Sujoy4 ,

Deo SVS5, Parshad Rajinder6, Ghosh Debabrata2, Sen Sudip1

1Dept. of Biochemistry,; 2Dept. of Physiology, 3Dept. of Pathology,; 4Dept. of GI Surgery, 5Dept. of

Surgical Oncology, 6Dept. Of General Surgery,

Presenting author: Aditi Bhattacharya, [email protected]

Corresponding author: Dr. Sudip Sen, [email protected]

Abstract Body Introduction: Cancer stem cells (CSCs) have been shown to be responsible for tumor

proliferation, metastasis and recurrence. Colorectal cancer, being asymptomatic in the earlier

stages, is already at advanced stage at the time of diagnosis.

Aim: Our study aims to identify molecular factors which are characteristic of colorectal cancer

stem cells and study if they can be developed as markers for a more accurate diagnosis and

prognosis.

Methodology: Stem cells were isolated from operative specimens of primary, untreated,

nonmetastatic, sporadic colorectal adenocarcinoma patients using documented markers antibodies

from tumor (CD44, CD166) and adjacent normal tissue (CD29, Lgr5) using fluorescence

activated cell sorting (FACS). RNA isolated from the sorted subsets were used to determine the

genome wide transcriptomic changes between CSC and normal stem cells (SC) by microarray.

Data was analysed using Flow Jo, Gene Spring GX13 and DAVID. Tumor sphere assay and

colony forming assay were done. Validation of putative gene targets identified by microarray was

done by RT-QPCR.

Results: The variability in CSC population (CD44+CD166+) was higher (range: 0.5-7%) than

normal SC (CD29+Lgr5+) (range:1-2%). The sorted CSC subset generated more tumor spheres

and had higher colony forming ability than normal SC subset. Microarray analysis comparing the

gene expression between CSC and normal SC RNA showed that both Wnt canonical and non-

canonical pathway molecules and intermediates were upregulated. Validation with real-time PCR

also showed that Wnt5a, Wnt2, Wisp1, TCF, LEF, Fzd10 and NfatC3 were upregulated in CSC

as compared to normal SCs.

Conclusion: Cancer stem cells have a greater sphere generating capacity and spheroids thus

formed are more proliferative in nature. The Wnt non-canonical pathway has been shown to play

a role in differentiation. Till date, very little research has implicated this pathway in

tumorigenesis and stemness. Our preliminary results have shown that the Wnt non-canonical pathway may contribute towards cancer stemness in colorectal cancer and studying this pathway

can help in developing better and more specific diagnostic and prognostic markers.


Phd-141

Using human fetal neural stem cells and oligodendrocytes as a disease model to delineate the

pathogenesis of cerebral palsy.

Dey D1*, Shrivastav V1, Bhat MA2, Singal C.M.S.4, Sharma JB3, Palanichamy JK1, Chattopadhyay P1,

Sinha S1, Seth P4, Sen S1

Presenting Author:

Name: Devanjan Dey

E-Mail: [email protected]


Name: Dr. Sudip Sen

E-Mail: [email protected]

Introduction:Cerebral Palsy (CP) is a neurological disorder in children. Oligodendrocytes (OL) are

vulnerable to hypoxic injury, resulting in CP. Fetal neural stem cells (FNSCs) can differentiate into

neuronal and glial lineages and “physiological hypoxic condition” influences their growth and

differentiation.

Aim and Objective:We aim to develop an in vitro model of CP by inducing FNSCs to differentiate into

OL, followed by exposure to hypoxia.

Materials and Methods:Aborted fetuses (n=4) were collected from the Department of Obstetrics

and Gynaecology, AIIMS, New Delhi, after Institute Ethics Committee clearance and informed

consent from mothers undergoing MTP. FNSCs were isolated from the sub-ventricular zone of the

fetal brain and expanded in culture. Immunocytochemistry (ICC) and flow cytometry were used to

evaluate Nestin and Sox2 expression in FNSCs. Annexin-V was used to analyze cell death. FNSCs were

exposed to different oxygen concentrations (20%, 6%, 2% and 0.2%) for 48 hours. Hypoxia exposure

was validated by qPCR for hypoxia markers. Microarray was done using Agilent whole genome 4x44K

array slides to study transcriptomic changes between FNSCs exposed to normoxia and hypoxia. Data

was analysed using Flow Jo, Gene Spring GX13 and GeneGO MetaCore.FNSCs were differentiated

into OL and evaluated for OL specific markers using ICC and qPCR.

Results and Conclusion: Nestin and Sox2 were found to be expressed by FNSCs by ICC and

flowcytometry. CA9, VEGF and PGK1 (hypoxia markers) were observed to be up regulated in FNSCs

exposed to hypoxia. Hypoxia did not result in cell death in FNSCs. As FNSCs differentiated into OL,

they expressed OL specific Myelinating Oligodendrocyte Glycoprotein (MOG) and NG-2 associated

with a decrease in Nestin and Sox-2 expression. Gene expression analysis of FNSCs exposed to

hypoxia revealed novel genes and some interesting pathways to be modulated. Validating

transcriptomic changes associated with hypoxia will help elucidate the pathogenesis of CP.



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