The need for an effective therapy for pancreatic ductal adenocarcinoma (PDAC) has prompted us to develop a novel combinatorial immunotherapeutic strategy for this very aggressive malignancy. In the design of our strategy we have been guided by the realization that to be effective, a therapy has to eradicate both differentiated PDAC cells and PDAC cancer initiating cells (CICs). According to the cancer stem cell theory, CICs play a major role in disease recurrence and metastatic spread, the major causes of patient morbidity and mortality. Furthermore, immunotherapy has to counteract the multiple escape mechanisms utilized by PDAC cells to avoid immune destruction. Therefore, i) to eliminate both differentiated PDAC cells and PDAC CICs, we have selected as a target B7-H3, since this tumor antigen (TA) can mediate the immune destruction of both types of targets. In addition, B7-H3 is up-regulated on tumor associated fibroblasts and stroma in the tumor microenvironment. Therefore, B7-H3 immunotargeting inhibits neoangiogenesis and contributes to the elimination of PDAC cells, even those with B7-H3 loss or down regulation. Lastly B7-H3 has a restricted distribution in normal tissues. Therefore, B7-H3 immunotargeting is not expected to cause side effects because of the targeting of normal tissues. ii) To counteract the escape mechanisms caused by abnormalities in HLA class I antigen processing machinery in PDAC cells, we have selected as effectors NK cells which do not require HLA class I antigen expression for recognition of target cells. To activate and expand the NK cell effector population at the level of the immune synapse, we have added IL-15 to the conventional bispecific NK Cell immune engagers (BiKEs) platform and used it to crosslink the scFv fragments derived from the B7-H3-specific mAb 376.96 to a highly modified camelid CD16 (FC?RIII)-specific scFv fragment. The latter binds to NK cells, while the former to PDAC cells. These novel hybrid molecules which are referred to as IL-15 TriKEs enhance the ability of NK cells to kill tumor cells with limited - if any - damage to normal cells. iii) To restore the susceptibility of PDAC cells to immune lysis we combine IL-15 TriKEs with the small molecule sonidegib, an inhibitor of the sonic hedgehog homolog pathway, since the activation of this pathway up-regulates the level of anti-apoptotic molecules. iv) To enhance the anti-tumor activity of NK cells we disrupt the PD-1/PD-L1 axis with an anti-PD-L1 mAb. We will test the hypothesis that NK cells in combination with IL-15 TriKEs, sonidegib and anti-PD-L1 mAb are effective in eradicating both differentiated PDACC cells and PDACC CICs, both in vitro and in vivo. As a result, PDAC disease recurrence and metastatic spread will be suppressed. Because of the limited budget the proposed studies will be performed only with PDAC cell lines. In future studies the potential clinical significance of the results obtained with cell lines will be assessed with patient derived xenografts. This information will then be translated to a clinical setting in a Phase I clinical trial.

Public Health Relevance

In response to the urgent need for effective therapies for the treatment of pancreatic ductal adenocarcinoma (PDAC), we have developed a novel combinatorial immunotherapeutic strategy which targets the tumor antigen B7-H3 utilizing IL-15 TtiKEs and NK cells. In addition, we counteract the hypoxia-induced escape mechanisms by inhibiting the Sonic Hedgehog Homologue pathway and by disrupting the PD-1/PD-L1 axis. We will test the ability of our combinatorial strategy to eradicate both differentiated PDAC cells and PDAC cancer initiating cells, both in vitro under hypoxia and in mice orthotopically grafted with PDAC cells. In addition, we will assess the toxicity caused by the administration of B7-H3 IL-15 TriKEs taking advantage of the crossreactivity of our human B7-H3-specific monoclonal antibody with its mouse counterpart. The information generated by the outlined studies will contribute to optimize the design of B7-H3 IL-15 TriKEs-based clinical trials for PDAC patients.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Small Research Grants (R03)
Project #
5R03CA231766-02
Application #
9741130
Study Section
Special Emphasis Panel (ZCA1)
Program Officer
Sommers, Connie L
Project Start
2018-08-01
Project End
2020-07-31
Budget Start
2019-08-01
Budget End
2020-07-31
Support Year
2
Fiscal Year
2019
Total Cost
Indirect Cost
Name
Massachusetts General Hospital
Department
Type
DUNS #
073130411
City
Boston
State
MA
Country
United States
Zip Code
02114
Pilla, Lorenzo; Ferrone, Soldano; Maccalli, Cristina (2018) Methods for improving the immunogenicity and efficacy of cancer vaccines. Expert Opin Biol Ther 18:765-784