Tumor-infiltrating lymphocyte (TIL) - Winship Cancer Institute of ...

Winship Regional Melanoma ConferenceAtlanta, GA February 9, 2013Tumor Infiltrating Lymphocyte (TIL)Based TherapyJeffrey WeberMoffitt Cancer CenterTampa, FL

Disclosures• I have consulted for BMS, Genentech and GSK for AdBoards and DSMBs• I have received less than $10,000 per year from anyof the above• I have served in the past on a Speaker’s Bureau forGenentech• My institution has received research funding fromBMS, Genentech and GSK

What Is Adoptive‐Cell Therapy?• T cells are isolated from tumorinfiltratinglymphocytes orperipheral blood• Ex vivo enrichment and expansionof antigenspecificeffector T cells utilizing– IL‐2, anti‐CD3, feedercells, and/or antigen‐pulsedDCsAdoptivetransferintopatientT cellsisolatedfrompatientIn vitroexpansionandactivation• T cells are reintroduced back to thepatientYee C. 2009 ASCO Educational Book. Alexandria, VA:American Society of Clinical Oncology. 2009;511‐515.

Tumor Infiltrating Lymphocyte(TIL) Therapy for melanoma• Adoptive cell therapy with TIL was developed in the 1980sbased on murine models at the NCI• Minced tumors were incubated with IL-2 ex vivo and it wasfound that over time the infiltrating lymphocytes grew outand cleared the tumor monolayer• The likelihood that cells would grow out was high (90%) inmelanoma, much less in other tumors• The cells were CD8 predominant, but occasionally a CD4+culture would grow• It took 4-5 weeks to expand the cells to 10 11 , and expansionwas performed in two stages:– Pre-expansion with just IL-2, then selection of reactive cells, and– Rapid expansion over 2 weeks with allo feeders and anti CD3

TIL cultures set-up n = 568Immediately or from previously cryopreserved sampleTIL grew?Y = 551Record of co-culture assay ?At any time prior to May 31 st , 2008NY89 Tumors censored(TIL frozen without testing)462 Tumors AnalyzedJin, J et al J Immunother 2012

ResultsSource of specimenSource of SpecimenSpecimen No. of tumors Percent positive P2ALL 462 58%Lymph node 152 50% 0.13Subcutaneous 190 54% 0.15Intraperitoneal 14 64% 0.79Liver 17 64% 0.8Lung 50 68% 0.18Bowel 13 54% 0.78• Overall 58% of the 462 tumors were able to generate reactiveTIL.• No significant differences between the sites of resection and theability to generate TIL.Jin, J et al J Immunother 2012

ResultsImpact of Prior Therapy within 3 months of harvestImpact of Prior Systemic TherapyTherapy No. of tumors Percent positive PSystemic therapy within 3 months of resection for TILIFNα-2b 3 100% 0.37Chemotherapy 11 45% 0.27Interleukin-2 79 70% 0.012Vaccine 7 42% 0.45Anti-CTLA4 antibody 1 67% 1.0Adoptive Cell Therapy 4 25% 0.31• IL‐2 administered within 3 months of TIL harvest resulted in a significantimprovement in the ability to generate tumor reactive TIL (70%p=0.012).Jin, J et al J Immunother 2012

Rapid Expansion of TIL forAdoptive Cell Therapy30-60 bags required!

APre-REPPerforin38.7CD8Perforin PerforinIL-2+4-1BB62.5CD8IL-245.4CD8BHernandez-Chacon et al. 2011 J ImmunotherCD8+Perf+ T-cell frequency (%)10090 P< 0.000180706050403020100Pre-REP IL-2 IL-2+41BBCDPre-REPCD8+GB+ T-cell frequency (%)1009080706050403020100P= 0.0077Pre-REP IL-2 IL-2+41BB

Is adoptive T cell therapy a promisingmelanoma therapy?• First TIL human clinical experience at NCI showed 11/20responses, but mostly PRs, mostly short lived responses, but inpreviously treated, bulky tumor; median OS< 12 mo• Trafficking experiments showed minimal tumor tropism, andtagging experiments showed rapid turnover• Subsequently, 29/86 patients treated with TIL + high dose IL‐2had a response; 85% got > 10 11 cells, no differences seen withprior IL‐2 or use of cytoxan• Large numbers of direct intra‐lesional TIL were not useful; whywere these non‐selected T cells not successful?• Response correlated with young TIL, and tumor specificityRosenberg et al, NEJM 1988, JNCI 1994

Lympho‐depletion: important role foradoptive T cell transfer• The use of non‐myeloablative chemotherapyprior to adoptive transfer of T cells in murinemodels allowed, in one maneuver, theelimination of T regulatory cells, the skewingof the T cell repertoire, and the promotion ofT cell memory by eliminating sinks forimportant cytokines like IL‐7 and IL‐15. Wouldlymphoid depletion prior to T cell adoptivetransfer prove useful?

Lymphoid depletion and ACT• Of 35 patients who failed high dose IL‐2 and receivedTIL after cytoxan+fludarabine preparative treatment toinduce lymphoid depletion, there was a 51% ORR, withmedian duration of response 12 months, with 3 CRs, 15PRs of 35• Subsequently, 43 patients had a similar preparativeregimen and different TIL growth technique: 49% ORRachieved• 25 additional patients each received the abovepreparative regimen, and either 2 or 12 cGy of TBI inaddition to chemotherapy; 54% and 72% ORR achievedwith only one treatment‐related death; most were priorhigh dose IL‐2 failuresDudley et al JCO 2005, 2008

Efficacy of TIL with lymphodepletion regimensRosenberg, S.A. et al; Clin Can Res 2011

Updated experience with TIL:the Surgery Branch NCIRosenberg, S.A. et al; Clin Can Res 2011

Survival by Prior Therapy In NCI‐SB“Selected” TIL ACT Trials•Progression on prior anti-CTLA4 does not preclude durable response•MD Anderson data: prior immunotherapy correlated with increased success for TILgeneration Rosenberg, S.A. et al; Clin Can Res 2011

Clinical images after adoptive T celltherapyRosenberg, SA and Dudley, M Curr Opin Immunol 2009

Adoptive cell therapy after lymphodepletion: increasedtelomere length in respondersDudley, M. E. et al. J Clin Oncol; 26:5233-5239 2008Copyright © American Society of Clinical Oncology

Other factors associated withoutcome with TILRosenberg, S.A. et al; Clin Can Res 2011

Waterfall plot of percentage change in tumor burden in all treated patients (n = 31):MD Anderson experienceRadvanyi L G et al. Clin Cancer Res 2012;18:6758-6770©2012 by American Association for Cancer Research

Comparison of total cells infused and major T-cell subsets in the infused TIL product betweenresponders and nonresponders: MD Anderson experinceRadvanyi L G et al. Clin Cancer Res 2012;18:6758-6770©2012 by American Association for Cancer Research

Comparison of PD-1, BTLA, and TIM-3 on CD8+ TIL in responders and nonresponders.Radvanyi L G et al. Clin Cancer Res 2012;18:6758-6770

Other TIL trials: Low dose IL‐2 and“Young TIL”• Ellebaek et al in J Trans Med 10:1 169 2012 treated 6patients with lymphodepletion, then TIL followed bylow dose IL2• 2 CR (30+, 10+ mos), 2 SD, 2 PD noted• Response associated with tumor reactive TIL• Besser et al in Clin Can Res 16: 2646 2010reported on 20 patients treated with unselected“young” TIL• 2 CR (18+, 4+), 8PR, 4 SD = 50% ORR• 90% success rate at treating patients with TIL• All responders had pre‐REP TIL by day 20

Outcome is associated with number of infused CD8 T cells:Danish experienceMarco Donia, advance online publication27 September 2012. doi:10.1038/jid.2012.336

PD-1 is predominantly expressed by tumor-infiltrating T cellscompared with peripheral blood T cells.Ahmadzadeh M et al. Blood 2009;114:1537-1544©2009 by American Society of Hematology

TcR transduced T cells for adoptive immunotherapy of cancerRosenberg, SA et al Nature Reviews Cancer 2008

Chimeric TcR transduced T cells for CLLPorter, DL et al N Engl J Med. 2011 August 25; 365(8): 725–733.

PD effect of adoptively transferred TcRtransduced cells: cytokine levelsPorter, DL et al N Engl J Med. 2011 August 25; 365(8): 725–733.

Combined antitumor activity of TCR engineered adoptive cell adoptive cell transfer (ACT)immunotherapy and vemurafenib in the pmel-1 model.Koya R C et al. Cancer Res 2012;72:3928-3937©2012 by American Association for Cancer Research

Conclusions• TIL are a promising therapy for stage IV melanoma, andbear testing in a randomized trial as second or thirdline therapy• Pre‐treatment with ipilimumab may augment thenumber and yield of TIL• Post‐transfer therapy with targeted drugs mayaugment the ability of TIL to mediate regression ofestablished tumor• TcR transduced TIL or PBMC can be very potenteffectors but represent a highly focused therapy towhich resistance may easily develop• TIL treated with anti CD137, and followed with PD‐1antibody may augment the yield, proliferation andactivity of cells after adoptive transfer

Acknowledgements• Bin Yu• WenShi Wang• Jay Alvarez• Ibrahim Younos• Amod Sarnaik• Erica Royster• Jim Mulé• Shari Pilon-Thomas