Non-viral precision TCR replacement
cancer, tumour
DISEASE PROGRESSION, SOLID TUMOUR, CANCER, DISEASE, TUMOUR, SEPARATION
Patients with relapsed or refractory metastatic solid tumours and the following criteria were considered for enrolment: disease progression with standard-of-care treatment options for their cancer; adequate performance status; and evidence of measurable disease and fulfilling eligibility criteria. These participants were considered for leukapheresis when up to three neoTCRs had been selected using the personalized prediction and isolation of neoTCRs from their PBMCs and tumour biopsy samples. Patients underwent leukapheresis at their local institution, and the leukapheresis product was shipped to the sponsor (PACT Pharma) for manufacture. CD4 and CD8 T cells were isolated using automated magnetic separation cell sorting and activated for 2 days. Next, cells were electroporated to introduce Cas9 protein, guide RNAs to knockout the endogenous The single-step precision genome-engineering process results in the replacement of the endogenous TCR with the patient’s native sequence neoTCR, the expression of which is placed under the regulation of the endogenous promoter (Fig. The potency of the final cell product was assessed through ELISAs to measure IFNγ secretion from neoTCR gene-edited T cells exposed to the cognate peptide–HLA multimer. The results were highly correlated (Pearson’s During the course of the clinical trial, the formulation of the culture medium was changed to improve cell growth and editing efficiency of the clinical-grade preparations (cell manufacture version 2.0 (v.2.0) to v.2.1), and we also changed the electroporation device (cell manufacture v.2.1 to v.3.0; Supplementary Table 