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Framework humanization increases the affinity and potency of anti-CD72 nanobody-based CAR-T cells for B-cell malignancies

ABSTRACT: Anti-CD19 CAR T cells can induce remissions in some patients with B-cell malignancies. However, new immunotherapeutic targets are urgently needed for the many who relapse. We recently described CD72 as a promising target in B-cell leukemia and lymphoma, developing fully synthetic nanobody-based CAR-T cells (nanoCARs) against this antigen. Toward clinical translation, here we humanize our previous nanobody framework regions, derived from llama, and surprisingly discover a clone ("H24") with enhanced potency against B-cell tumors both in vitro and in vivo. In vitro, H24 nanoCARs showed improved cytokine secretion and favorable immunophenotypic properties. RNA sequencing before and after tumor exposure reveals humanized H24 CD72 nanoCARs have unique transcriptional programs compared to CD19 CAR T cells. We further find that H24 nanoCARs lead to sustained CD72 downregulation on tumors treated in vivo. Underpinning this improved potency, H24 had higher binding affinity to CD72 compared to a fully llama framework. Further affinity maturation moderately increased cytotoxicity versus antigen-low tumors. This work supports clinical translation of H24 CD72 nanoCARs, reveals potential mechanisms of resistance to this cellular therapy, and unexpectedly demonstrates that nanoCAR potency can be improved by framework alterations alone, without any change to the complementarity determining regions (CDRs) of the nanobody sequence.

ORGANISM(S): Homo sapiens

PROVIDER: GSE218791 | GEO | 2022/12/29

REPOSITORIES: GEO

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