Leaders in Pharmaceutical Business Intelligence (LPBI) Group

LIVE 9/19 4PM – 5:30PM NK CELL-BASED CANCER IMMUNOTHERAPY @CHI’s 14th Discovery On Target, 9/19 – 9/22/2016, Westin Boston Waterfront, Boston

 

 

CHI’s 14th Discovery On Target, 9/19 – 9/22/2016, Westin Boston Waterfront, Boston

http://www.discoveryontarget.com/

http://www.discoveryontarget.com/crispr-therapies/

#BostonDOT16

@BostonDOT

Leaders in Pharmaceutical Business Intelligence (LPBI) Group is a Media Partner of CHI for CHI’s 14^th Annual Discovery on Target taking place September 19 – 22, 2016 in Boston.

In Attendance, streaming LIVE using Social Media

Aviva Lev-Ari, PhD, RN

Editor-in-Chief

http://pharmaceuticalintelligence.com

COMMENTS BY Stephen J Williams, PhD

Technological Innovations enabling NK Cell based Cancer immunotherapies

 

4 PM Autologous ex-vivo expanded NK cells for Solid Tumor Immunotherapies

Ali Ashkar DVM PhD McMaster university

 

NK cells

1. Healer reg NK cells
2. Inflammatory NK like TAMS
3. Cytotoxic 90% in blood but in healthy tissue not that much

 

CD56+ CD16- healer NK near fetal maternal interface

 

Ex vivo expansion from breastCA donor most wind up cd56+ they produce IFN gamma

Their model is using ascites and testing in a PDX model but that is immunosuppressed, what about immunocompetent

 

Her2+ cell lines are extremely sensitive what about her2- then? Or triple negative?

 

Do NK cells from lung kill lung cancer cells? Yes both stimulated and unstimulated NK cells specifically

 

Ovarian ascites NK cells can purify and expand and produce cytokine

However they used survival as endpoint so possibly not reflective of NK effect on tumor

 

Other problem with model have to give them IL2 or IL15 or human NK cells would not survive in mice; however humanized mice might work better as don’t need exogenous IL2 or IL15 (mice produce the humanized version of these

 

4:30 Novel CARs introduced into NK cells facilitate Potent Tumor Cell Killing

 

NK-92 based CAR-tumor targeting (TNK) CEA CARTNK

 

CD19 CARTNK  CD123 CARTNK CD38 CAR-TNK

 

The CEA CAR work on colon cancer cell lines and all CEA positive lines however there is a NK resistant LS174T cell line

 

Early killing using a CAR NK don’t see with just NK and introducing a CAR provides killing for resiistant tumors

So the CD18 CARTNK worked well on Raji cells

 

Used the CD123 CARTNK on RPMI 8226 cell line so it would appear that you have to personalize the CAR NK cell model with the markers on each tumor – what about tumor heterogeneity?

In Raji they get about a 50% improvement in tumor killing using a CAR

There is significantly higher CD38 expression in the tumor setting

 

5) Understanding of NK Cell Effector Functions: A Single Cell Lab on a Chip Perspective

 

Tania Konry Northeastern University

 

Dynamics of cancer cell signaling responses to immune cell signals

 

High throughput single cell characterization of the effects of immune cell heterogeneity

 

Phenotypic drug profiling for better targeting of drug-resistant tumors

 

So with a microfluidic can encapsulate a cell with an assay/reagent

Could you encapsulte with nanoparticle?

 

They hold IP for a bead based multiplex assay.  The microfluidic system is a platform to detect heterogeneity among tumor cells.

 

Looking at interaction of T-cell tumor cell and NK cell using multiplex bead assays in a microfluidic system

 

Can detect cell killing within the droplet in the microfluidic system and test different drugs as well as cell death kinetics

For Example they had looked at the interaction of PD-L1 with the NK cell and tumor cell by coencapsulating the tumor and effector cells

 

With breast cancer cell spheroids you can use alginate as a polymer to generate those spheres and can perfuse with drug of your choice

 

TECHNOLOGICAL INNOVATIONS ENABLING NK CELL-BASED CANCER IMMUNOTHERAPY

4:00 Autologous ex vivo Expanded NK Cells for Solid Tumor Immunotherapy

Ali Ashkar, D.V.M., Ph.D., Professor, Pathology and Molecular Medicine, McMaster Immunology Research Centre, McMaster University

In healthy individuals, the innate immune system, particularly natural killer (NK) cells, are crucial for immune surveillance. In patients with advanced tumors number and activity of NK cells decline significantly. Recent advances in NK cell expansion and activation have generated renewed interest in adoptive NK cell therapy for cancers. We have expanded NK cells from blood of breast, lung and ovarian cancer patients and have investigated their activities against autologous primary tumor cells. In addition, we have established xenograft models with the primary tumors to study the antitumor activities of autologous NK cells against primary tumor cells in vivo. Ex vivo expanded NK cells survive and proliferate in vivo in the presence of autologous PBMCs.

• CD56 and CD16 – phynotype in Human
• Healer NK cells (regNK)
• Inflammatory NK cells (Similar to TAM)
• In Malignant Ascites – NK cells CD16 negative – changed phynotype from positive
• Model of autologous NK Cell CAncer Immunotherapy for OC, BC and LC
• Cytokine Activated Cancer patients vs Health donors
• Expanded NK cells IV control group and NK injected
• Stimulated vs unstimulated NK cells: All cells, non-Epithelial, Epithelial cells
• NK treated and IL-2 + NK Treated vs COntrol (IL-12): Liver vs Lung
• Ex vivo expanded autologous PB-NK vs allogenous PB-NK – NK from ovarian cancer patients: Peripheral blood vs Ascites
• Expand NK to produce IFN-gamma
• Can expanded-NK cells from ascites kill aotologous primary ovarian cancer cells in vivo? Untreated vs NK treated
• CD3 vs CD56 – in vivo expansion of NK cells in the peritoneal cavity?

4:30 Novel CARs Introduced into NK Cells Facilitate Potent Tumor Cell Killing that Results in Tumor Regression

Rohit Duggal, Ph.D., Director, Experimental Cellular Therapy, Sorrento Therapeutics

This presentation features an introduction of chimeric antigen receptors (CARs), which provide homing and specificity to cytotoxic cells of the immune system. Novel CARs isolated from Sorrento’s G-MAB library targeting various tumor antigens will be described. The characterization of the NK cells modified to express these CARs will also be described.

• NK Cell therapy

1. mmodification od existing NK tumor cells engineered
2. stem-cells

• Proof of Concept:

1. NK-92 antiCEA – CD28z 4inf  – specificity – colon cancer carcinoma : NK-92 vs Anti CEA
2. improve potency of CAN upon expression of the anti CEA CAR in NK-resistant
3. STI-NK 0n targeting NK (TNK)
4. tumor control in orthotopic colon cancer model
5. Tumor regression in Colon Cancer upon treatment with anti CEA
6. NK-resistant LS74T colon carcinoma cells

• GBM patient segmentation by TCGA

1. STI NK line cytotoxicity against K562 LEUKEMIA LINE
2. CAR-TNK
3. Anti CD38 CAR-TNK killing specific to CD38 expressing cells – low killing

Conclution

• NK Products in Tumor models
• Tumorigenic capabilities of NK and of STI NK

 

5:00 Understanding of NK Cell Effector Functions: A Single-Cell Lab-on-a-Chip Perspective

Tania Konry, Ph.D., Assistant Professor, Department of Pharmaceutical Sciences, Northeastern University

Natural Killer (NK) cells are an essential component of innate immunity that actively inhibit tumor development. Here we present a novel single-cell method of analyzing the mechanisms underlying the cellular interactions of NK cells with multiple myeloma cells. The integrated droplet microfluidics device developed by our group permits compartmentalization of cell pairs and secreted products within sub-nanoliter volumes and thereby controls cell-to-cell communication by limiting it to interactions between the co-encapsulated cells. It allows monitoring of both contact-dependent (immune synapse formation, delivery of lytic hits) and contact-independent cellular interactions (release of cytokines, chemokines) simultaneously. This dynamic single-cell experimental model is expected to provide preclinical information particularly relevant to the scenario of NK cell-cancer cell interactions.

• DC- T cell
• Antigen on DC-T interaction
• DC-T-Tumor cell interaction
• cytotoxic molecule – secretion
• Beads
• modeling Myeloma Cell
• Dynamic analysis of NK-tumor cell interaction: cell motility >> conjugation >> detachment >> lytic hit >> cell death
• Brefeldin inhibits cytokine secretion – in presence of NK cells: FAst vs slow kill
• Cancer cell lysis: EGTA blocks calcium-dependent perforin polymerization
• Duration od contact, killing time: contact dependent and Independent and Target cell death: Response to PD-L1 blockage
• Immunotherapy regulation
• conjugate duration, detection of lytic hit and cell death

Conclusion

1. Novel 3D Tumor mimicking microenvironment to differentiate aggressive breast cancer from Indolent Cancers and to evaluate cancer Drugs
2. LAB-on-a CHIP: 3D Tumor cell model

DFCI/NEW Joint Program in Cancer Drug Development

 

5:30 Close of Symposium