Exploiting tumour infiltrating lymphocytes (TILs) as a therapeutic strategy in epithelial ovarian cancer

Transcription

1 Exploiting tumour infiltrating lymphocytes (TILs) as a therapeutic strategy in epithelial ovarian cancer G.L.Owens 1,2, V.Sheard 2, M.Price 1, D.E.Gilham 2, R.J.Edmondson 1 1 Gynaecological Oncology Research Group, Institute of Cancer Sciences, University of Manchester, UK 2 Clinical and Experimental Immunotherapy Group, Institute of Cancer Sciences, Manchester Cancer Research Centre, University of Manchester, UK

2 HGSOC - Typical clinical course Maintenance? TIL Therapy Bowtell et al., Nature Reviews Cancer 215; 15:

3 Ovarian cancer is immunogenic Monoclonal antibodies Cancer vaccines Adoptive cell therapy Cancer antigen Dendritic cell Effector cell Cancer cell NK cells NK Gamma delta Immune escape mechanisms Zhang et al., N Engl J Med 23; 348: Immune checkpoint inhibitors (PD-1/PD-L1)

4 Adoptive cell Adoptive therapy TILs therapy nnnnnnn isolated In vitro activation and expansion of tumour-specific Pre-conditioning nnnnnnnnnn nnnnnnn Tumour excised (1) Natural anti-tumour Rapid Chemotherapy expansion of tumour-specific and/or Expansion of radiotherapy for 2-3 weeks Identify reactive Identify reactive Tumour-specific returned to patient +/- Cytokines

5 Aims Project aims 1 1. Test the reproducibility of an established protocol to expand TILs from EOC biopsies. 2. Define the anti-tumour function, specificity and phenotype of EOC TILs. 3. Determine the impact of cryopreservation on the functional activity of TILs.

6 Experimental Adoptive Approach TILs therapy nnnnnnn Tumour nnnnnnnnnn Disaggregation nnnnnnn of tumour Co-culture with autologous tumour and IFN gamma ELISA (1) Natural anti-tumour Cells mitogenically stimulated using anti-cd3/-cd28 magnetic beads + IL-2 nnn n Expansion of for 2-3 weeks Identify reactive nnnnn nnnn NNNNNN NNNNN Phenotype by flow cytometry

7 TILs can readily Isolation be expanded & Expansion from of EOC TILs A Primary B IDS N u m b e r o f T IL s x N u m b e r o f T IL s x D a y s a fte r is o la tio n D a y s a fte r is o la tio n C N u m b e r o f T IL s x 1 6 (D a y 1 9 ) ID S N S P r im a r y TILs expanded from 19/21 (>9%) biopsies No difference re. expansion from primary and IDS biopsies

8 Ovarian TILs show features associated with central & effector memory phenotypes 1 8 Naïve CD27 + CCR7 + CD62L + CD45RA + % L y m p h o c y te s Central memory CD27 + CCR7 + CD62L + CD45RA - Effector memory CD27 -/low CCR7 -/low CD62L - CD45RA - Effector CD27 - CCR7 - CD62L - CD45RA + C D 4 C D % C D 4 + L y m p h o c y te s % C D 8 + L y m p h o c y te s C D 2 7 C D 2 8 C D 4 5 R A C D 4 5 R O C D 2 5 C D 6 2 L C C R 7 C D 2 7 C D 2 8 C D 4 5 R A C D 4 5 R O C D 2 5 C D 6 2 L C C R 7

9 Ovarian TILs retain functional activity against autologous tumour cells 5 5 *** T IL o n ly 5 T IL + T u m o u r p g /m l IF N g pg/ml IFNy *** ** ** * ** * * ** ** *** ** * ** ** * 5 M O C 6 M O C 7 M O C 2 7 M O C 2 9 M O C 3 M O C 3 1 S o lid M O C 3 1 A s c ite s M O C 3 3 M O C 4 1 M O C 3 4 M O C 3 6 M O C 3 7 M O C 3 8 M O C 4 M O C 4 2 M O C 4 3 M O C 4 6 Mean and SD of three replicates are shown. *P <.5, **P <.5, ***P <.5.

10 Ovarian TILs retain functional activity following cryopreservation M O C 2 7 fre e z e -th a w M O C 2 9 fre e z e -th a w M O C 3 1 fre e z e -th a w p g /m l IF N g ** ** p g /m l IF N g ** **** p g /m l IF N g ** * F r e s h T IL F r e s h T IL + T u m o u r T h a w e d T IL T h a w e d T IL + T u m o u r F r e s h T IL F r e s h T IL + T u m o u r T h a w e d T IL T h a w e d T IL + T u m o u r F r e s h T IL F r e s h T IL + T u m o u r T h a w e d T IL T h a w e d T IL + T u m o u r M O C 3 3 fre e z e -th a w M O C 3 4 fre e z e -th a w M O C 3 6 fre e z e -th a w *** * p g /m l IF N g * * p g /m l IF N g 5 * ** p g /m l IF N g F r e s h T IL F r e s h T IL + T u m o u r F r e s h T IL + A s c ite s T h a w e d T IL T h a w e d T IL + T u m o u r T h a w e d T IL + A s c ite s F r e s h T IL F r e s h T IL + T u m o u r T h a w e d T IL T h a w e d T IL + T u m o u r F r e s h T IL F r e s h T IL + T u m o u r T h a w e d T IL T h a w e d T IL + T u m o u r Mean and SD of three replicates are shown. *P <.5, **P <.5, ***P <.5, ****P <.1.

11 Summary What s next TILs can be readily expanded from EOC biopsies Expanded TILs retain functional activity against autologous tumour Expanded TILs demonstrate less differentiated phenotypes Indicates TILs likely to engraft and persistent in circulation, increasing effector response Future work: Working toward early phase clinical trial Molecular profiling of EOC tumours and TCRs of TILs

12 Acknowledgments Supervisory team Prof Richard Edmondson Dr David Gilham Dr Jaime Honeychurch Clinical and Experimental Immunotherapy Group Vicky Sheard Dr Antje Schutt Dr Gray Kueberuwa Dr Milena Kalaitsidou Dr Manon Pillai Dr Vanessa Clay Aoife Kilgallon Adam Milner Abdulrahman Bahashwan Weiming Zheng Funding Cancer Research UK, Manchester Centre Gynaecological Oncology Research Group Marcus Price Clinical Gynaecological Oncology team Dr Rick Clayton Dr Cath Holland Dr Emma Crosbie Mr Saad Ali St Mary s Hospital theatre and clinic staff Patients CMFT Biobank Dr Jay Brown Core Facilities, Paterson Institute Jeff Barry Garry Ashton

13 Proposed Adoptive clinical trial TILs flowchart therapy Expand for 2-3 weeks Reactive cryopreserved Rapid expansion Collect T-cells from Tumour Biopsy (1) Natural anti-tumour Primary or IDS surgery bbc c bbc c Completion of chemotherapy Return cells to patient + supportive therapy Identify reactive

14 Stratification Adoptive of patients TILs therapy I M M U N O S C O (1) R Natural anti-tumour E High Immunoscore Intermediate Immunoscore Low Immunoscore Adoptive cell therapy, Immune checkpoint blockade Agents which stimulate CD8 T cell trafficking & infiltration Chimeric antigen receptor (CAR) Identify reactive

15 In vivo Dynabeads

16 Early ACT Adoptive trials in ovarian TILs therapy cancer Author, year Regime Pa1ents Outcomes Aoki, TILs expanded in IL-2 - Infused IV following cyclophosphamide - TILs only vs TILs + chemo Freedman, 1994 (1) Natural anti-tumour - TILs expanded in IL-2 - Infused IP with high dose IL-2 - IL-2 vs TILs + IL-2 Fujita, TILs expanded with an[- CD3 and IL-2 - Infused IV following comple[on of chemo - No TILs vs TILs Identify reactive N = 17 Stage III/IV or recurrent EOC N=11 Stage III/IV, all chemo-resistant N = 24 Stage II, III or IV, with no detectable lesion following primary treatment CR of 14.3% TILs only cohort & 7% TILs + chemo cohort No objec[ve response 3-year OS: 1% in TILs cohort, 67.5% in controls

17 PD-1 Adoptive TILs therapy (1) Natural anti-tumour Identify reactive