Antigen specific anti-SPAS-1 CD8 T cell responses to in ... · standard local treatments. •Best...
Transcript of Antigen specific anti-SPAS-1 CD8 T cell responses to in ... · standard local treatments. •Best...
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Antigen specific anti-SPAS-1 CD8 T cell responses to in situ vaccination using
irreversible electroporation (IRE) of prostate tumors
Joseph R. [email protected]
M.D. Candidate, Class of 2020University of Minnesota Medical School, Twin Cities
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Prostate Cancer• Radical prostatectomy, external-beam radiotherapy, and brachytherapy are all
standard local treatments.• Best data for the efficacy and safety of local treatment are from two randomized
trials that compared radical prostatectomy against watchful waiting.✓ Surgery was associated with greater urinary incontinence and erectile
dysfunction.
Attard, G., Parker, C., Eeles, R. A., Schröder, F., Tomlins, S. A., Tannock, I., … De Bono, J. S. (2016). Prostate cancer. The Lancet, 387(10013), 70–82. http://doi.org/10.1016/S0140-6736(14)61947-4
• Disease in many patients recurs after local therapy.✓ Optimum timing to start systemic treatment initiation is not clear.✓ The best treatment of the prostatic primary tumor in patients with
metastatic disease at diagnosis is also unclear.
• Novel treatment modalities:✓ immunotherapy✓ irreversible electroporation
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Immunotherapy
http://newsroom.cumc.columbia.edu/cancer-immunotherapy/
(anti-CTLA-4)
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Tumor lysis and antigen release by irreversible electroporation (IRE)
Waitz, R., Hou, Y., Rim, T., Greenberg, N. M., Shastri, N., Fong, L., & Allison, J. P. (2008). specific T cells )/ SH3GLB2 : A prostate tumor antigen identified by CTLA-4 blockade, 105(9), 3509–3514.
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SPAS-1 release from TRAMP C2 tumor cells by irreversible electroporation (IRE)
Waitz, R., Hou, Y., Rim, T., Greenberg, N. M., Shastri, N., Fong, L., & Allison, J. P. (2008). specific T cells )/ SH3GLB2 : A prostate tumor antigen identified by CTLA-4 blockade, 105(9), 3509–3514.
• First T cell-defined transgenic adenocarcinoma of mouse prostate (TRAMP) tumor antigen.
• Spas-1 ortholog SH3GLB2 is overexpressed in prostate cancer metastases.
• SH3 DOMAIN, GRB2-LIKE protein.
• SPAS-1 epitope SNC9-H8 arose from a point mutation in one allele of the gene in TRAMP tumor cells.
• Immunization with dendritic cells pulsed with SNC9-H8 peptide resulted in protection against TRAMP-C2 tumor challenge.
• MHC-I tetramers are available
Tramp C2 Tumor Cell
Tumor Antigen (SPAS-1)
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Hypothesis:IRE combined with anti-CTLA4 immunotherapy will produce more SPAS-1 specific T cells and SPAS-1 specific T memory cells in vivo versus conventional resection with anti-CTLA4 antibody treatment.
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Inject 106 Tramp C2 Tumor Cells
Allow tumor growth for specified number of days
CD8 T Cell Response to 1o Tumor and Spas-1 Tetramer
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Inject 106 Tramp C2 Tumor Cells
Allow tumor growth for 14 days
Treat with resection or IRE with or without anti-CTLA4 antibody
Treat with or without anti-CTLA4 antibody via IP injection
Experiment Scheme
Sacrifice mouse and harvest spleen and lymph nodes
Use antibody tetramers to enrich for lymphocytes and SPAS1 T Cells
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Gating Scheme for SPAS-1 Specific T CellsSS
C-A
FSC-A
SSC-
A
SSC-W Live/Dead
Thy
1.2
CD8
Beta
CD8 alpha
SPAS
-1 T
etra
mer
SPAS-1 Tetramer
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Figure 1. Total SPAS-1 specific T CD8 T cells 11 days after resection or IRE with or without immunotherapy.
Treatment
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Figure 1. Total SPAS-1 specific CD8 T cells 11 days after resection or IRE with or without immunotherapy.
Treatment
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Figure 2. Total SPAS-1 specific T CD8 T cells 31 days after resection or IRE with or without immunotherapy.
Treatment
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Conclusions
• IRE combined with anti-CTLA4 immunotherapy results in greater total numbers of SPAS-1 specific CD8 T cells versus resection with anti-CTLA4 antibody. • IRE may provide an alternative means to resection of prostatic
disease due to preservation of local tissue, potentially decreasing the risk of fistula, urinary incontinence, and erectile dysfunction.• IRE has potential for host vaccination to treat metastatic disease and
recurrence.
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Future Directions
•Anti-CTLA4 titration studies to eliminate potential immunopathologies. •Treat with anti-CTLA4 antibody sooner (day 0, 3, 6, and 9 instead of
day 1, 4,7, 10)•Enroll mice with tumor volumes ideal for resection by IRE.•Enroll more mice •Apply IRE to different mouse model•Treat mice with only anti-CTLA4 antibody•Assess T cell function with IFNɣ and TNFα
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Acknowledgments
Thanks to :The Shimizu Lab
• Dr. Burbach• Dr. Shimizu• Buck O’Flanagan• Katharine Young
• UMN Medical School• UMN Center for Immunology• UMN Medical Student Summer Research Program in Infection and
Immunity• UMN Dept. of Biomedical Engineering• NIH T35 Research Training Grant
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Figure 3. Percentage of mice tumor free following treatment and secondary tumor challenge.
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Cryoablation with anti-CTLA-4 therapy
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Cryoablation with anti-CTLA-4 therapy
Norton, L., & Allison, J. P. (2015). cryoablation with anti-CTLA-4 therapy, 72(2), 430–439. http://doi.org/10.1158/0008-5472.CAN-11-1782.Potent
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Spas-1 Specific T Cell Function 18 Days Post 2o Tumor Challenge
IFNg
TNFa
Sple
en
IgG
IgG
Anti-CTLA4
Anti-CTLA4
Tum
or
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CTV
CD44
Buck O’Flanagan BO047; BO054 ff First Batch, Ind. Expts
A Trp2 T cells + DCs
No Antigen
+ Trp2 Peptide10ng/ml
+ B16 Cryocycle(total lysis)1.2 x 105 cell equiv
+ B16 IRE lysate
1.2 x 105 cell equiv
In vitro DC-mediated Trp2 T cell activation by B16 lysates: IRE > Cryo > Heat
CB
B16 Lysate (Cell Equivalents)
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