192 - A Novel Therapeutic Approach Using ENO1-specific CAR-NK Cells to Target KRAS-Mutated Cancers
Presenter(s)
K. S. C. Chao1, C. Huang2, H. Y. Chang3, W. C. Huang4, T. T. Yuan4, Y. Y. Tsai2, Y. S. Lin2, J. A. Liang5, and K. C. Y. Huang6; 1China Medical University Hospital, China Medical University, Taiwan, Taiwan, 2China Medical University Hospital, Taichung, Taiwan, 3National Yang Ming Chiao Tung University, HsinChu, Taiwan, 4HuniLife Biotechnology, Taipei, Taiwan, 5Department of Radiation Oncology, China Medical University Hospital, Taichung City, Taiwan, 6China Medical University, Taichung, Taiwan
Purpose/Objective(s): KRAS mutation occurs in multiple cancers types (82% PDAC, 38% CRC and 22% NSCLC, etc.), and due to its intranuclear nature, currently the therapeutic options are very limited. In addition, KRAS mutated cancer is also resistant to radiotherapy. We have previously reported that these cells tend to be in the state of high immunometabolism and upregulate invasive and metastatic activity via cell surface translocation of a moonlighting glycolysis protein, ENO1. Here, we present a novel approach using chimeric antigen receptor (CAR)-NK cells targeting membrane-bond ENO1 and evaluate the combination effect with radiotherapy in KRAS-mutated cancers.
Materials/Methods: We first examined the relationship between KRAS status, surface ENO1 and immune cell infiltration in PDAC (n=150), CRC patients (n=180), and NSCL (n=180) to determine the correlation of surface ENO1 and immune cell profiles based on the KRAS status. We further developed ENO1-specific CAR-NK cells with a novel therapeutic ENO1 antibody (HuL001). We then assess the antitumor efficacy of ENO1 CAR-NK against KRAS-mutated cancer cells with varying levels of surface ENO1 expression, both in vitro and using tumor-bearing syngeneic mice in combination with radiotherapy.
Results: We found that surface expression of ENO1 was upregulated in KRAS mutation and also causing immunosuppressive TME. ENO1-targeted CAR-NK cells demonstrated target-specific cytotoxicity when compared to naive NK cells. CRISPR/Cas9 mediated knockout of ENO1 in target cells further confirmed the specific killing ability of ENO1 CAR-NK cells. ENO1 CAR-NK cells significantly increased the response to low-dose radiotherapy in vitro. Moreover, ENO1 CAR-NK cells enhanced the therapeutic response of local radiotherapy in vivo, showing superior tumor inhibition and survival time compared to the RT-only group in KRAS-mutated tumor-bearing mice.
Conclusion: These results showed that ENO1 served as an ideal surrogate cell surface target for KRAS mutated cancers. ENO1-specific CAR-NK when in combination with RT overcome the resistant nature of KRAS-mutated cancers. This approach can be applied to patients who currently have very limited therapeutic options.