Investigating anti-Glypican 2 (GPC2) chimeric antigen receptor (CAR) T-cell trafficking and tumor microenvironment (TME) interaction in fully immunocompetent small cell lung cancer (SCLC) mouse models
Final Report Abstract
CAR T-cell manufacturing utilizing CRISPR/Cas9 is an exciting avenue that has the potential to increase both the access to CAR T-cells and accelerate translation of novel CAR constructs due to ease of good manufacturing practice (GMP) grade template DNA production. However, the non-viral manufacture of CAR T-cells is associated with challenges resulting from modest insertion rates and low target cell numbers respectively. During my postdoctoral fellowship I have identified a novel insertion strategy to engineer T-cell via CRISPR knock-in (HITI) and optimized the conventional insertion method to enable the manufacture of non-viral CRISPR knock-in CAR T-cells. In addition, I applied CRISPR knock-in CAR T-cells against B7-H3 in small cell lung cancer (SCLC), and here identified that low-antigen SCLC as well as non-neuroendocrine SCLC, which shows an epithelial-to-mesenchymal cell state with high levels of TGFb, can be efficiently addressed with B7-H3 CAR T-cells co-expressing cJUN. Finally, I have provided evidence that the manufacture of non-viral engineered T-cells (cJUN+B7-H3 CAR) with large transgenes (3.6kb) inserted in a targeted manner is feasible at clinical scale.
Publications
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Homology-independent targeted insertion (HITI) enables guided CAR knock-in and efficient clinical scale CAR-T cell manufacturing. Molecular Cancer, 22(1).
Balke-Want, Hyatt; Keerthi, Vimal; Gkitsas, Nikolaos; Mancini, Andrew G.; Kurgan, Gavin L.; Fowler, Carley; Xu, Peng; Liu, Xikun; Asano, Kyle; Patel, Sunny; Fisher, Christopher J.; Brown, Annie K.; Tunuguntla, Ramya H.; Patel, Shabnum; Sotillo, Elena; Mackall, Crystal L. & Feldman, Steven A.
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Non-viral chimeric antigen receptor (CAR) T cells going viral. Immuno-Oncology and Technology, 18, 100375.
Balke-Want, H.; Keerthi, V.; Cadinanos-Garai, A.; Fowler, C.; Gkitsas, N.; Brown, A.K.; Tunuguntla, R.; Abou-el-Enein, M. & Feldman, S.A.
