Targeting the intracellular immune checkpoint CISH with CRISPR-Cas9-edited T cells in patients with metastatic colorectal cancer: a first-in-human, single-centre, phase 1 trial

Emil Lou¹, Modassir S Choudhry², Timothy K Starr³, Timothy D Folsom⁴, Jason Bell⁴, Blaine Rathmann⁴, Anthony P DeFeo⁴, Jihyun Kim⁴, Nicholas Slipek⁴, Zhaohui Jin⁵, Darin Sumstad⁶, Christopher A Klebanoff⁷, Katherine Ladner⁸, Akshat Sarkari⁸, R Scott McIvor⁹, Thomas A Murray¹⁰, Jeffrey S Miller¹¹, Madhuri Rao¹², Eric Jensen¹², Jacob Ankeny¹², Mahmoud A Khalifa¹³, Anil Chauhan¹⁴, Benjamin Spilseth¹⁵, Ajay Dixit¹⁶, Paolo P Provenzano¹⁷, Wenjing Pan¹⁸, Daniel Weber¹⁸, Miranda Byrne-Steele¹⁸, Tom Henley², David H McKenna¹⁹, Matthew J Johnson⁴, Beau R Webber⁴, Branden S Moriarity⁴

Affiliations

¹ Department of Medicine, Division of Hematology, Oncology, and Transplantation, University of Minnesota, Minneapolis, MN, USA. Electronic address: emil-lou@umn.edu.
² Intima Bioscience, New York, NY, USA.
³ Department of Obstetrics, Gynecology, and Women's Health, University of Minnesota, Minneapolis, MN, USA.
⁴ Department of Pediatrics, University of Minnesota, Minneapolis, MN, USA; Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA; Center for Genome Engineering, University of Minnesota, Minneapolis, MN, USA.
⁵ Deparment of Medical Oncology, Mayo Clinic, Rochester, MN, USA.
⁶ Cell Therapy Clinical Laboratory, University of Minnesota Medical School, Minneapolis, MN, USA.
⁷ Department of Medicine, Human Oncology and Pathogenesis Program, and Parker Institute for Cancer Immunotherapy, Memorial Sloan Kettering Cancer Center, New York, NY, USA.
⁸ Department of Medicine, Division of Hematology, Oncology, and Transplantation, University of Minnesota, Minneapolis, MN, USA.
⁹ Department of Genetics, Cell Biology, and Development, University of Minnesota, Minneapolis, MN, USA.
¹⁰ Division of Biostatistics and Health Data Science, University of Minnesota Medical School, Minneapolis, MN, USA.
¹¹ Department of Medicine, Division of Hematology, Oncology, and Transplantation, University of Minnesota, Minneapolis, MN, USA; Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA.
¹² Department of Surgery, University of Minnesota Medical School, Minneapolis, MN, USA.
¹³ Department of Laboratory Medicine and Pathology, University of Minnesota Medical School, Minneapolis, MN, USA.
¹⁴ Department of Radiology, University of Kansas Medical Center, Kansas City, KS, USA.
¹⁵ Department of Radiology, University of Minnesota Medical School, Minneapolis, MN, USA.
¹⁶ Department of Biomedical Engineering, University of Minnesota, Minneapolis, MN, USA; Division of Gastroenterology, Hepatology, and Nutrition, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USA.
¹⁷ Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA; Department of Biomedical Engineering, University of Minnesota, Minneapolis, MN, USA; Center for Multiparametric Imaging of Tumor Immune Microenvironments, University of Minnesota, Minneapolis, MN, USA; Stem Cell Institute, University of Minnesota, Minneapolis, MN, USA.
¹⁸ iRepertoire, Huntsville, AL, USA.
¹⁹ Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA; Cell Therapy Clinical Laboratory, University of Minnesota Medical School, Minneapolis, MN, USA.

PMID: 40315882
DOI: 10.1016/S1470-2045(25)00083-X

Clinical Trial

Targeting the intracellular immune checkpoint CISH with CRISPR-Cas9-edited T cells in patients with metastatic colorectal cancer: a first-in-human, single-centre, phase 1 trial

Emil Lou et al. Lancet Oncol. 2025 May.

. 2025 May;26(5):559-570.

doi: 10.1016/S1470-2045(25)00083-X. Epub 2025 Apr 29.

Authors

Affiliations

¹ Department of Medicine, Division of Hematology, Oncology, and Transplantation, University of Minnesota, Minneapolis, MN, USA. Electronic address: emil-lou@umn.edu.
² Intima Bioscience, New York, NY, USA.
³ Department of Obstetrics, Gynecology, and Women's Health, University of Minnesota, Minneapolis, MN, USA.
⁴ Department of Pediatrics, University of Minnesota, Minneapolis, MN, USA; Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA; Center for Genome Engineering, University of Minnesota, Minneapolis, MN, USA.
⁵ Deparment of Medical Oncology, Mayo Clinic, Rochester, MN, USA.
⁶ Cell Therapy Clinical Laboratory, University of Minnesota Medical School, Minneapolis, MN, USA.
⁷ Department of Medicine, Human Oncology and Pathogenesis Program, and Parker Institute for Cancer Immunotherapy, Memorial Sloan Kettering Cancer Center, New York, NY, USA.
⁸ Department of Medicine, Division of Hematology, Oncology, and Transplantation, University of Minnesota, Minneapolis, MN, USA.
⁹ Department of Genetics, Cell Biology, and Development, University of Minnesota, Minneapolis, MN, USA.
¹⁰ Division of Biostatistics and Health Data Science, University of Minnesota Medical School, Minneapolis, MN, USA.
¹¹ Department of Medicine, Division of Hematology, Oncology, and Transplantation, University of Minnesota, Minneapolis, MN, USA; Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA.
¹² Department of Surgery, University of Minnesota Medical School, Minneapolis, MN, USA.
¹³ Department of Laboratory Medicine and Pathology, University of Minnesota Medical School, Minneapolis, MN, USA.
¹⁴ Department of Radiology, University of Kansas Medical Center, Kansas City, KS, USA.
¹⁵ Department of Radiology, University of Minnesota Medical School, Minneapolis, MN, USA.
¹⁶ Department of Biomedical Engineering, University of Minnesota, Minneapolis, MN, USA; Division of Gastroenterology, Hepatology, and Nutrition, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USA.
¹⁷ Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA; Department of Biomedical Engineering, University of Minnesota, Minneapolis, MN, USA; Center for Multiparametric Imaging of Tumor Immune Microenvironments, University of Minnesota, Minneapolis, MN, USA; Stem Cell Institute, University of Minnesota, Minneapolis, MN, USA.
¹⁸ iRepertoire, Huntsville, AL, USA.
¹⁹ Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA; Cell Therapy Clinical Laboratory, University of Minnesota Medical School, Minneapolis, MN, USA.

PMID: 40315882
DOI: 10.1016/S1470-2045(25)00083-X

Erratum in

Correction to Lancet Oncol 2025; 26: 559-70.
[No authors listed] [No authors listed] Lancet Oncol. 2025 Jun;26(6):e298. doi: 10.1016/S1470-2045(25)00273-6. Lancet Oncol. 2025. PMID: 40449503 No abstract available.

Abstract

Background: Over the past decade, immunotherapeutic strategies-mainly targeting the PD-1-PD-L1 immune checkpoint axis-have altered cancer treatment for many solid tumours, but few patients with gastrointestinal forms of cancer have benefited to date. There remains an urgent need to extend immunotherapy efficacy to more patients while addressing resistance to current immune checkpoint inhibitors. The aim of this study was to determine the safety and anti-tumour activity of knockout of CISH, which encodes cytokine-inducible SH2-containing protein, a novel intracellular immune checkpoint target and a founding member of the SOCS family of E3-ligases, using tumour infiltrating lymphocyte (TILs) genetically edited with CRISPR-Cas9 in patients with metastatic gastrointestinal epithelial cancers.

Methods: For this first-in-human, single-centre, phase 1 trial, patients aged 18-70 years with a diagnosis of metastatic gastrointestinal epithelial cancer with progressive disease following at least one first line standard therapy, measurable disease with at least one lesion identified as resectable for TIL generation and at least one other lesion meeting RECIST criteria as measurable to serve as an indicator of disease response, and an ECOG performance status of 0 or 1 were screened and enrolled if meeting these and all other eligibility criteria. TILs procured from tumour biopsies were expanded on the basis of neoantigen reactivity, subjected to CRISPR-Cas9-mediated CISH knockout, and infused intravenously into 12 patients after non-myeloablative lymphocyte depleting chemotherapy (cyclophosphamide 60 mg/kg per dose on study days -6 and -5, and fludarabine 25 mg/m² per dose on days -7 to -3) followed by high-dose IL-2 (aldesleukin; 720 000 IU/kg per dose). The primary endpoint was safety of administration of neoantigen-reactive TILs with knockout of the CISH gene, and a key secondary endpoint was anti-tumour activity measured as objective radiographic response and progression-free and overall survival. This study is registered with ClinicalTrials.gov, NCT04426669, and is complete.

Findings: Between May 12, 2020, and Sept 16, 2022, 22 participants were enrolled in the trial (one patient was enrolled twice owing to lack of TIL outgrowth on the first attempt); ten patients were female, and 11 were male (self-defined). One patient was Asian, the remainder were White (self-defined). We successfully manufactured CISH knockout TIL products for 19 (86%) of the patients, of whom 12 (63%) received autologous CISH knockout TIL infusion. The median follow-up time for the study was 129 days (IQR 15-283). All 12 (100%) patients had treatment-related severe adverse events. The most common grade 3-4 adverse events included haematological events (12 patients [100%]) attributable to the preparative lymphodepleting chemotherapy regimen or expected effects of IL-2, fatigue (four patients [33%]), and anorexia (three patients [25%]). Deaths of any cause for patients on study were attributed to the underlying disease under study (metastatic gastrointestinal cancer) and related complications (10 patients) or infection (grade 5 septicaemia in one patient). There were no severe (≥grade 3) cytokine release or neurotoxicity events. Six (50%) of 12 patients had stable disease by day 28, and four (33%) had stable disease ongoing at 56 days. One young adult patient with microsatellite-instability-high colorectal cancer refractory to anti-PD1/CTLA-4 therapies had a complete and ongoing response (>21 months).

Interpretation: These results support the safety and potential antitumour activity of inhibiting the immune checkpoint CISH through the administration of neoantigen-reactive CISH-knockout TILs, with implications for patients with advanced metastatic cancers refractory to checkpoint inhibitor immunotherapies, and provide the first evidence that a novel intracellular checkpoint can be targeted with therapeutic effect.

Funding: Intima Bioscience.

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Conflict of interest statement

Declaration of interests BSM and BRW have received Sponsored Research Agreement funding from Intima Bioscience that funded portions of this work. EL reports the following disclosures: equipment for laboratory-based research 2018 to present from Novocure, honorarium for panel discussion organised by Antidote Education for a CME module on diagnostics and treatment of HER2-positive gastric and colorectal cancers, funded by Daiichi-Sankyo, compensation for scientific review of proposed printed content from Elsevier Publishing and Johns Hopkins Press, consultant for Nomocan Pharmaceuticals (no financial compensation), financial interests in Ryght, present or past research funding as Institutional Principal Investigator for clinical trials sponsored by Celgene, Novocure, Intima Bioscience, and the National Cancer Institute, and University of Minnesota membership in the Caris Life Sciences Precision Oncology Alliance (no financial compensation), and present or past volunteer for Medical Advisory Boards for patient advocacy groups Colon Cancer Coalition, Minnesota Colorectal Cancer Research Fund, Gilda's Club-Twin Cities (no financial compensation). All other authors declare no competing interests.

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Targeting the intracellular immune checkpoint CISH with CRISPR-Cas9-edited T cells in patients with metastatic colorectal cancer: a first-in-human, single-centre, phase 1 trial

Affiliations

Targeting the intracellular immune checkpoint CISH with CRISPR-Cas9-edited T cells in patients with metastatic colorectal cancer: a first-in-human, single-centre, phase 1 trial

Authors

Affiliations

Erratum in

Abstract

Conflict of interest statement

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Associated data

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Full Text Sources

Medical

Research Materials

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