作者
Alessia Potenza,Chiara Balestrieri,Martina Spiga,Luca Albarello,Federica Pedica,Francesco Manfredi,Beatrice Claudia Cianciotti,Claudia de Lalla,Oronza A. Botrugno,Cristina Faccani,Lorena Stasi,Elena Tassi,Silvia Bonfiglio,Giulia Maria Scotti,Miriam Redegalli,Donatella Biancolini,Barbara Camisa,Elena Tiziano,Camilla Sirini,Monica Casucci,Chiara Iozzi,Danilo Abbati,Fabio Simeoni,Dejan Lazarevič,Ugo Elmore,Guido Fiorentini,Giulia Di Lullo,Giulia Casorati,Claudio Doglioni,Giovanni Tonon,Paolo Dellabona,Riccardo Rosati,Luca Aldrighetti,Eliana Ruggiero,Chiara Bonini
摘要
Colorectal tumours are often densely infiltrated by immune cells that have a role in surveillance and modulation of tumour progression but are burdened by immunosuppressive signals, which might vary from primary to metastatic stages. Here, we deployed a multidimensional approach to unravel the T-cell functional landscape in primary colorectal cancers (CRC) and liver metastases, and genome editing tools to develop CRC-specific engineered T cells.We paired high-dimensional flow cytometry, RNA sequencing and immunohistochemistry to describe the functional phenotype of T cells from healthy and neoplastic tissue of patients with primary and metastatic CRC and we applied lentiviral vectors (LV) and CRISPR/Cas9 genome editing technologies to develop CRC-specific cellular products.We found that T cells are mainly localised at the front edge and that tumor-infiltrating T cells co-express multiple inhibitory receptors, which largely differ from primary to metastatic sites. Our data highlighted CD39 as the major driver of exhaustion in both primary and metastatic colorectal tumours. We thus simultaneously redirected T-cell specificity employing a novel T-cell receptor targeting HER-2 and disrupted the endogenous TCR genes (TCR editing (TCRED)) and the CD39 encoding gene (ENTPD1), thus generating TCREDENTPD1KOHER-2-redirected lymphocytes. We showed that the absence of CD39 confers to HER-2-specific T cells a functional advantage in eliminating HER-2+ patient-derived organoids in vitro and in vivo.HER-2-specific CD39 disrupted engineered T cells are promising advanced medicinal products for primary and metastatic CRC.