GRC NPC – Rooney, Cliona

Cliona Rooney (Baylor College of Medicine, USA)
“Cancer Immunotherapy and Challenges for NPC”

1. T cell for each patient, pre existing , 22% chance response
2. Vaccine an Ad-based LMP2, MVA-EBNA1-LMP2
3. LCL activated vEBVST
4. Singpore TESSA therapeutics (http://goo.gl/zdbYpA) 330 patients (與Wed Poster #5相關)
5. Ad LMP1/2/LCL activated T cell for NPC, not better for EBVST, but is better in lymphoma patients (type 2 malignancy lymphoma)
6. SFC, IFNG coated plate, 6 month 1.5 year
7. survivors epitome spread: MAGEA4 + SURVIVIN + PRAME
8. Dendritic cells are difficult to make in NPC patients
9. 1st stimulation: EBV EBNA1 ,LMP1, LMP2, BARF1
10. 2nd stimulation: Pepmix all possible MHC2
11. GRACE trial (patients of lymphoma)
12. Switch to IL15/7
13. High dose IL15 increase specificity
14. Fewer CD4 cells in high dose IL15 (SF: IL21 and IL15 are part of the gamma-chain cytokine family and are crucial for survival and proliferation of Tfh (follicular Th), cytotoxic and memory T cells, both IL21 and IL15 are being used in clinical trial)
15. IL15H increases central key EBVSTs
16. Post infusion expansion of EBVSTs in patients with lymphoma
17. heterogeneity in NPC TME
18. Importance for epitope spreading, maybe viral lytic genes are important (Mei-Ru asked what kind of lytic genes in her mind, she replied: IE genes)
19. TGFBR
20. Infiltrating barrier
21. Gottschalk PI is responsible
22. Lympho depletion/Reactivate EBV
23. Immuno checkpoint
24. Genetic TGFBR DN to increase T cells
25. Steve Rosenberg was mentioned again and again and again (https://goo.gl/LKXqYG)
26. T cells express NPC expressing cytokine 2 receptor to “homing”
27. GuangZhou published some promising results (ref: Oncoimmunology 2015)

From Ming-Han 

1. 1 Small molecules? (one drug for all patients)
2. 2 EBV-specific T cells;
3. 3 Chimeric antigen receptor (CAR) T cells?
4. 4 Gene-modified T cells? (EBVSTs)
5. One patient one drugs?

Ongoing trials

1. Chiun Hsu (許駿); NTU PD1 inhibitor (KEYTZUDA KEYNOTE 028 Overall Response rate 10.8 %)
2. LMP2 peptide pulsed DCs (HK): increase LMP2A specific T cells
3. Ad-dLMP1-LMP2 transduced DCs (Singapore by Prof. Toh): DTH but no elevated LMP specific T cells.
4. MVA-EBNA1-LMP2: functional in 50% patients with the elevated T cell responses.
5. LCL-activated EBVSTs for NPC (adjuvant (62%) remain diseases free / 2nd or relapse in 48.7%)
6. LCL activated EBVSTs after Chemotherapy? (Singapore (Toh, Ho): 35 patients; 97% type III diseases; Gemicitabin and carboplatin 4 cycles. (combine treatment?); 71.4% responses; overall survival 2yr 65%; 5yr 35%.)
7. Gemicibin alone vs Gemicibin + EBVSTs on going in 350 patients. (Ongoing)

Ad5f35-∆LMP1-LMP2 (Adenovim)

• infect PBMC –> boosted by EBV-LCLs (in between plus IL2) —> wish to have LMP2A/1 T cell CTLs. (Bollard et al, J, Clin Onco 2014 32 (8) 798-808.)

EBVST treat for EBV lymphoma (CTC project)

to NKT lymphoma (remission 25/26)
– Epitope spreading after treatment of EBVSTs seems very important.

This strategy seems good BUT problems

• too long
• No LCL available from these EBV+lymphoma patients; either low B cell or the B cells can’t be immortalised by EBV for?? reason.
• Require live virus.
• Many cases all T cells only in the end recognise EBNA3C.

Original plan:

• (PBMC infected by B95-8) –> LCL –> stimulated with DCs with LMP1/2A ––> T cells (peptide of LMP2A by using many many peptides; each 15mer but with 11aa overlapping each)

Now: 

• PBMC+peptide –> (+IL4/7) can have expansion 100X times T cells only within 10 days; enough time to treat PTLD.

Issue:

• This strategy works super well in healthy donor BUT IL4/7 doesn’t work well for expansion of patient’s T cell for ?? reason.

Phase I clinical trial design (GRACE)

• HL, BL… EBV+ lymphoma.
• Patients T cells with very low avidity;
• Patient with lymphoma have very few T cells.
• Growth of T cells from patients always badly.

New strategy:

IL4/7 vs IL7/15? which is better: 
found IL7/15 is better (high dose IL15: 100ng/ml; medium dose IL15: ? ng/ml)
But still IL7/15 still difficult to expand T cells from patients.

             IL4+IL7       IL7+IL15     IL7+IL15hi
CD4          50%          30%                ~0
CD8          50%          70%             ~100 %

Problem2: LMP1/2 actually not expressed in all tumor cells. For example; if stimulate T cells with BARF1, LMP2, EBNA1 or LMP1, in the end T cells only recognise one peptides…

Epitope spreading: 

• SSX2, NYESP, Survivin, Prame, Mage4 —> these antigens only exists in lymphoma.
• after treatment patients have T cells against these peptides then the treatments outcome will be better.
• So far: 2/10 cases total cure; 4/10 with better response.

For NPC cases: 

• EBVSTs have less responses for NPC patients
• Possible reasons:  solid tumor; difficult for T cells to enter; Immune suppressive environment.

Requirements if want to treat NPC:

• migration to tumor site
• Infiltrate tumor parenchtmas?
• have to proliferate at tumor sites that express also MHCI

@Gottschalk (NPC clinical trials)
@Effective CAR and TIL stimulation
Requirement for human trials: GMP director; CMP CTL; everything GMP

——————————–Q1: CD4/8 mix during T cell expansion; which one is better?