Development of HER2-Specific Canine Chimeric Antigen Receptor-Natural Killer Cells for Canine Caner Immunotherapy — ASN Events
  1. Schedule
  2. Poster Session One
  3. Development of HER2-Specific Canine Chimeric Antigen Receptor-Natural Killer Cells for Canine Caner Immunotherapy

Development of HER2-Specific Canine Chimeric Antigen Receptor-Natural Killer Cells for Canine Caner Immunotherapy (#150)

Geum-Seop Kim 1 2 , Yurim An 3 , Jae-Hee Park 1 , Duck Cho 4 , Je-jung Lee 5 , Sang-ki Kim 1 2 3
  1. Department of Companion and Laboratory Animal Science, College of Industrial Science, Kongju National University, Yesan-gun, Chungnam, Republic of Korea
  2. Research Institute for Natural Products, Kongju National University, Yesan-gun, Chungnam, Republic of Korea
  3. Department of Applied Biotechnology, Kongju National University, Yesan-gun, Chungnam, Republic of Korea
  4. Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea
  5. Department of Hemotology-Oncology, Chonnam National Univresity Hwasun Hospital, Hwasun, Jeollanamdo, Republic of Korea

Canine mammary tumors (CMTs) exhibit substantial parallels with human breast cancer, including tumor progression pathways, molecular traits, and clinical features, making them valuable models in comparative oncology. This research explores a novel immunotherapy strategy utilizing chimeric antigen receptor natural killer (CAR-NK) cells for treating HER2 expressing cancer in dogs. We engineered an anti-HER2 CAR specifically targeting HER2-positive canine cancers and incorporated it into canine NK cells, aiming to develop an advanced and effective therapeutic approach. To enhance lentiviral transduction efficiency, we optimized packaging conditions by testing different ratios of the transfer plasmid, psPAX2, and envelope vector. The optimized ratio of the transfer plasmid, psPAX2, and envelope vector resulted in the highest transduction efficiency. Additionally, envelope receptor expression analysis of ASCT-1, ASCT-2, and LDLR on canine NK cells was conducted to determine the optimal transduction conditions. Flow cytometry analysis confirmed a significant enrichment of NK cells after expansion, with the proportion of non-B, non-T NK lymphocytes rising from 13.6% in PBMCs to 79.1% by day 11, while T and B cell populations declined accordingly. Successful transduction of canine CAR-NK cells revealed 24.4% CAR+ cells, with stable expression maintained up to 11 days post-transduction. Crucially, canine CAR-NK cells exhibited significantly enhanced cytotoxicity against both canine mammary tumor cells (CMT-U334, CF41.mg) and human breast cancer cells (SKBR-3) compared to non-transduced NK cells, as confirmed by CCK-8 and RTCA assays. This cytotoxic effect was dose-dependent and consistently observed across multiple cell lines. This study lays a strong foundation for innovative immune cell therapy in canine cancer treatment while also offering potential translational insights for human breast cancer therapies. It underscores the versatility and efficacy of CAR-NK cells as a groundbreaking therapeutic approach in oncology.