Developing TCR gene therapy for multiple myeloma
description
Transcript of Developing TCR gene therapy for multiple myeloma
![Page 1: Developing TCR gene therapy for multiple myeloma](https://reader034.fdocuments.in/reader034/viewer/2022051700/568167f4550346895ddd6bf1/html5/thumbnails/1.jpg)
Developing TCR gene therapy for multiple myeloma
Gavin BendleLLR Bennett Senior FellowSchool of Cancer SciencesUniversity of Birmingham
![Page 2: Developing TCR gene therapy for multiple myeloma](https://reader034.fdocuments.in/reader034/viewer/2022051700/568167f4550346895ddd6bf1/html5/thumbnails/2.jpg)
•Introduction to TCR gene therapy of cancer
•Assessing the value of TCR gene therapy in a autochthonous mouse cancer model
•Pre-clinical development of TCR gene therapy of MM
Overview
![Page 3: Developing TCR gene therapy for multiple myeloma](https://reader034.fdocuments.in/reader034/viewer/2022051700/568167f4550346895ddd6bf1/html5/thumbnails/3.jpg)
Multiple myeloma
•Plasma cell malignancy that is the second most common haematological malignancy worldwide
•Novel agents (e.g. lenalidomide, bortezomib) have contributed to increase in average survival times from 4 to 8 years in the last decade
• Despite these advances MM remains largely incurable
Need for new therapeutic approaches that not only increase survival times but are also ultimately curative
![Page 4: Developing TCR gene therapy for multiple myeloma](https://reader034.fdocuments.in/reader034/viewer/2022051700/568167f4550346895ddd6bf1/html5/thumbnails/4.jpg)
• T cell immunity to tumours can be induced by adoptive T cell therapy:
• Transfer of T cells
• T cell replete allogeneic-HSCT & DLI for haematological malignancies• Mortality/morbidity due to GVHD limits use
• Transfer of genes encoding the TCR
• Ag-specificity of a T cell determined by TCR• Endow patient T cells with tumour-reactivity of a defined Ag-specificity• Destroy tumours without damaging normal tissues
Adoptive T cell therapy
![Page 5: Developing TCR gene therapy for multiple myeloma](https://reader034.fdocuments.in/reader034/viewer/2022051700/568167f4550346895ddd6bf1/html5/thumbnails/5.jpg)
Cancer patient
Removal of peripheralblood lymphocytes
Infusion of autologous TCR gene modified T cells
Ex vivo transduction process
TCR gene therapy
• TCR gene transfer is conceptually attractive:– Generate large numbers of defined antigen-specific patient T cells– Generate tumor-reactive specificities not present in pre-existing T cell repertoire
![Page 6: Developing TCR gene therapy for multiple myeloma](https://reader034.fdocuments.in/reader034/viewer/2022051700/568167f4550346895ddd6bf1/html5/thumbnails/6.jpg)
CT scan of liver metastasis in patienttreated with TCR-modified T cells
Clinical trials shown feasibility and potential of TCR gene therapy
![Page 7: Developing TCR gene therapy for multiple myeloma](https://reader034.fdocuments.in/reader034/viewer/2022051700/568167f4550346895ddd6bf1/html5/thumbnails/7.jpg)
Clinical testing of TCR gene therapy for MM has commenced
![Page 8: Developing TCR gene therapy for multiple myeloma](https://reader034.fdocuments.in/reader034/viewer/2022051700/568167f4550346895ddd6bf1/html5/thumbnails/8.jpg)
Safety risks associated with TCR gene therapy
Expression profile of target antigen in normal tissues: the critical parameter determining the safety of TCR gene therapy
• Most targets of TCR gene therapy are tumor-associated self-antigens
• Toxicity may occur if target antigen is also expressed by some normal tissues
• If vital normal tissues express target antigen toxicity can be severe or even fatal
![Page 9: Developing TCR gene therapy for multiple myeloma](https://reader034.fdocuments.in/reader034/viewer/2022051700/568167f4550346895ddd6bf1/html5/thumbnails/9.jpg)
Additional genetic modification of T cells as a strategy to enhance TCR gene therapy efficacy
![Page 10: Developing TCR gene therapy for multiple myeloma](https://reader034.fdocuments.in/reader034/viewer/2022051700/568167f4550346895ddd6bf1/html5/thumbnails/10.jpg)
Can additional genetic modification of TCR transduced T cells be used to obtain durable clinical responses with TCR gene therapy?
• TCR gene transfer endows patient T cells with desired antigen-specificity
• Can additional genetic modification endow these cells with optimal functional properties?
Used a mouse model of prostate carcinoma to assess if additional genetic modification of T cells enhances TCR gene therapy efficacy
![Page 11: Developing TCR gene therapy for multiple myeloma](https://reader034.fdocuments.in/reader034/viewer/2022051700/568167f4550346895ddd6bf1/html5/thumbnails/11.jpg)
TRAMP mice: prostate tumour model
TRansgenic Adenocarcinoma of the Mouse Prostate(Greenberg et al, PNAS 1995)
SV40 large T antigen under control of a rat probasin promoter
expression Large T
intraepithelial
neoplasia (PIN)
Microinvasivecarcinoma
3 8-12 16
Invasivecarcinoma
24
![Page 12: Developing TCR gene therapy for multiple myeloma](https://reader034.fdocuments.in/reader034/viewer/2022051700/568167f4550346895ddd6bf1/html5/thumbnails/12.jpg)
• Block by co-transducing TCR Td T cells with dnTGFRII
• Rationale for blockade of TGF- signalling in TCR transduced T cells:
• TGF- signalling inhibits CTL proliferation & effector functions
• Elevated TGF-expression in human malignancies including prostate cancer
• Elevated levels of TGF- in prostate of TRAMP mice with advanced prostate cancer
Does TGF- signalling blockade in TCR transduced T cells promote tumour regression in TRAMP mice?
![Page 13: Developing TCR gene therapy for multiple myeloma](https://reader034.fdocuments.in/reader034/viewer/2022051700/568167f4550346895ddd6bf1/html5/thumbnails/13.jpg)
Group I: Pre-conditioning (5Gy TBI) + non Td T cells
Group II: Pre-conditioning (5Gy TBI) + SV40 TCR Td T cells
Group III: Pre-conditioning (5Gy TBI) + dnTGFRII Td T cells
Group IV: Pre-conditioning (5Gy TBI) + SV40 TCR & dnTGFRII co-Td T cells
Does blockade of TGF- signalling in TCR transduced T cells lead to tumour regression in TRAMP mice?
expression Large T
intraepithelial
neoplasia (PIN)
Microinvasivecarcinoma
3 8-12 16
Invasivecarcinoma
24
Week 24 28
Histopathology
![Page 14: Developing TCR gene therapy for multiple myeloma](https://reader034.fdocuments.in/reader034/viewer/2022051700/568167f4550346895ddd6bf1/html5/thumbnails/14.jpg)
Regression of advanced prostate cancer in TRAMP mice after TCR gene therapy and blockade of TGF- signalling
![Page 15: Developing TCR gene therapy for multiple myeloma](https://reader034.fdocuments.in/reader034/viewer/2022051700/568167f4550346895ddd6bf1/html5/thumbnails/15.jpg)
Tumour regression in TRAMP mice after TCR gene therapy & TGF- blockade
Non-Td SV40 TCR TdSV40 TCR &
dnTGFRII co-Td
SV40 Large T Ag Immunostaining
TGF-1Immunostaining
Ki67Immunostaining
Is the tumour regression observed at 28 wks of age sustained?
![Page 16: Developing TCR gene therapy for multiple myeloma](https://reader034.fdocuments.in/reader034/viewer/2022051700/568167f4550346895ddd6bf1/html5/thumbnails/16.jpg)
Sustained regression of advanced prostate cancer in TRAMP mice after TCR gene therapy and blockade of TGF- signalling
Does the observed tumor regression lead to enhanced survival?
![Page 17: Developing TCR gene therapy for multiple myeloma](https://reader034.fdocuments.in/reader034/viewer/2022051700/568167f4550346895ddd6bf1/html5/thumbnails/17.jpg)
Enhanced survival of TRAMP mice after TCR gene therapy & TGF- blockade
![Page 18: Developing TCR gene therapy for multiple myeloma](https://reader034.fdocuments.in/reader034/viewer/2022051700/568167f4550346895ddd6bf1/html5/thumbnails/18.jpg)
• Blockade of TGF- signalling in TCR transduced T cells promotes tumour regression in TRAMP mice with advanced prostate cancer
• Demonstrates potential of additional genetic modification of TCR transduced T cells to enhance TCR gene therapy efficacy
• Elevated expression of TGF- in many human malignancies including MM suggest that this approach warrants clinical testing
Summary
![Page 19: Developing TCR gene therapy for multiple myeloma](https://reader034.fdocuments.in/reader034/viewer/2022051700/568167f4550346895ddd6bf1/html5/thumbnails/19.jpg)
Pre-clinical development ofTCR gene therapy for MM
![Page 20: Developing TCR gene therapy for multiple myeloma](https://reader034.fdocuments.in/reader034/viewer/2022051700/568167f4550346895ddd6bf1/html5/thumbnails/20.jpg)
TCR α-chain
TCR β-chain
TCR a- & -chains are generated by genetic rearrangement
• Sequence information in TCR - & -chain genes that defines TCR specificity:– V element used– J element used– Nucleotide sequence at the junction between V-J elements
• Cloning of TCR genes is a bottleneck in production of new TCRs– Developed high throughput strategy to isolate TCR gene sequences for TCR gene therapy
![Page 21: Developing TCR gene therapy for multiple myeloma](https://reader034.fdocuments.in/reader034/viewer/2022051700/568167f4550346895ddd6bf1/html5/thumbnails/21.jpg)
TCR gene capture technology
Shear gDNA & enrich for gDNA fragments
encoding TCR & loci
Paired-end deep sequencing
Align sequence pairs to reference genome to identify
rearrangements at TCR & loci
Reconstruction of TCR - & -chain genesIdentify potentially clinically relevant T cell specificities
in biological material using multiplexing technology
FACS sort single T cells of desired specificity,expand in vitro for 14 days and extract gDNA
Linnemann et al. Nature Med in press
![Page 22: Developing TCR gene therapy for multiple myeloma](https://reader034.fdocuments.in/reader034/viewer/2022051700/568167f4550346895ddd6bf1/html5/thumbnails/22.jpg)
A library of cancer-testis antigen-specific TCRs assembled using TCR gene capture
Rapidly assembled a panel of tumor-reactive TCRs with the potentialto be used for TCR gene therapy of a variety of malignancies
Target antigen Restriction Peptide # TCRs Material
MAGE-C2 A2 ALKDVEERV 2 PBMC
MAGE-C2 A2 LLFGLALIEV 1 TIL
MAGE-C2 A2 KVLEFLAKL 3 PBMC
MAGE-A10 A2 GLYDGMEHL 2 TIL
SSX-2 A2 MLAVISCAV 3 PBMC
MAGE A2 A2 YLQLVFGIEV 3 PBMC
NY-ESO-1 A2 SLLMWITQA 1 PBMC
LAGE-1 A2 MLMAQEALAFL 1 TIL
MAGE-A1 B7 RVRFFFPSL 1 TIL
HERV-Kmel A2 MLAVISCAV 2 TIL
TAG A3 RLSNRLLLR 2 TIL
![Page 23: Developing TCR gene therapy for multiple myeloma](https://reader034.fdocuments.in/reader034/viewer/2022051700/568167f4550346895ddd6bf1/html5/thumbnails/23.jpg)
Cancer-testis antigens
•Cancer-testis (C/T) antigens:
• Expressed in human germ line & variety of human malignancies
• Attractive targets for TCR gene therapy of MM:
• Expression in MM cells in a high frequency of patients• Examples:
• MAGE C1: ~ 70%• MAGE C2: ~ 50%
• Absent from normal tissues accessible to immune system
![Page 24: Developing TCR gene therapy for multiple myeloma](https://reader034.fdocuments.in/reader034/viewer/2022051700/568167f4550346895ddd6bf1/html5/thumbnails/24.jpg)
Validation and characterisation of C/T Ag-specific TCRs
•Assess which C/T Ag-specific TCRs are best suited to clinical translation of TCR gene therapy for MM:
– C/T Ag expression frequency in MM patient population– Efficacy & safety of T cells transduced with different C/T Ag-specific TCRs
C/T Ag-specific TCRs isolated to date
Target antigen Restriction Peptide # TCRs Material
MAGE-C2 A2 ALKDVEERV 2 PBMC
MAGE-C2 A2 LLFGLALIEV 1 TIL MAGE-C2 A2 KVLEFLAKL 3 PBMCMAGE-A10 A2 GLYDGMEHL 2 TIL
SSX-2 A2 MLAVISCAV 3 PBMCMAGE A2 A2 YLQLVFGIEV 3 PBMCNY-ESO-1 A2 SLLMWITQA 1 PBMC
LAGE-1 A2 MLMAQEALAFL 1 TILMAGE-A1 B7 RVRFFFPSL 1 TIL
TAG A3 RLSNRLLLR 2 TIL
![Page 25: Developing TCR gene therapy for multiple myeloma](https://reader034.fdocuments.in/reader034/viewer/2022051700/568167f4550346895ddd6bf1/html5/thumbnails/25.jpg)
Blockade of TGF- signalling in C/T Ag-specific TCR transduced T cells
•Additional genetic modification of TCR transduced T cells to tailor their activity can enhance TCR gene therapy efficacy
•TGF- in MM microenvironment leads to myeloma-induced T cell dysregulation
•Selectively block TGF- signalling in C/T Ag-specific TCR transduced T cells
– Generate and validate a retroviral construct encoding the relevant C/T Ag-specific TCR and a dominant-negative TGF- receptor
Primary benefit: novel clinical trials of TCR gene therapy in MM patients in B’ham
![Page 26: Developing TCR gene therapy for multiple myeloma](https://reader034.fdocuments.in/reader034/viewer/2022051700/568167f4550346895ddd6bf1/html5/thumbnails/26.jpg)
•TCR gene therapy holds promise as a treatment for cancer
•Additional genetic modification of TCR transduced T cells to tailor their activity and enhance therapeutic efficacy
•TCR gene capture utilized to assemble a library of C/T Ag-specific TCRs that can be used for TCR gene therapy of various cancers
•Assessing which TCRs are best suited to clinical translation of TCR gene therapy for multiple myeloma
•Pre-clinical development will be followed by clinical testing in B’ham
Summary
![Page 27: Developing TCR gene therapy for multiple myeloma](https://reader034.fdocuments.in/reader034/viewer/2022051700/568167f4550346895ddd6bf1/html5/thumbnails/27.jpg)
Acknowledgements
•NKI, Amsterdam– Carsten Linnemann– Laura Bies– Kaspar Bresser– Bianca Heemskerk– Pia Kvistborg– Roel Kluin– Ron Kerkhoven– Marja Nieuwland– Ji-Ying Song– John Haanen– Ton Schumacher
•Shemyakin & Ovchinnikov Institute, Moscow
– Dmitriy Chudakov– Dmitriy Bolotin
•MDC, Berlin– Xioajing Chen– Thomas Blankenstein
•Cancer Sciences, B’ham– Guy Pratt– Paul Moss