Universal CAR-T Cells (REVO-UWD-01) for Metastatic Colorectal Cancer

This is an investigator initiated trial to assess the efficacy and safety of a GCC-targeting CAR-T therapy (REVO-UWD-01) in the metastatic colorectal cancer. It also aims to explore the feasibility of using a novel universal CAR-T cell platform.

The study will use T cells from healthy donors, modified using a novel universal CAR-T technology, to treat metastatic colorectal cancer patients. The antigen-binding site of the CAR molecule recognizes GCC as the target.

The main questions it aims to answer are:

* What is the maximum tolerated dose (MTD) of GCC-CAR-T therapy in universal CAR-T cell treatments?
* What are the dose-limiting toxicities (DLT) and treatment-emergent adverse events (TEAE)?
* What is the treatment\&amp;amp;#39;s efficacy, as measured by objective response rate (ORR) and progression-free survival (PFS)? Researchers will assess whether universal CAR-T cells have good safety and efficacy in treating colorectal cancer, while improving accessibility and lowering treatment costs.

Participants will:

* Receive universal GCC-CAR-T cells through a 3+3 dose escalation scheme.
* Undergo chemotherapy conditioning before CAR-T infusion.
* Be monitored for adverse events, immune response, and disease progression.

The study will collect data on both short-term outcomes (within the first few months post-treatment) and long-term safety and efficacy.

Eligibility Criteria:
Inclusion Criteria:

1. Age: 18-80 years old;
2. Pathologically confirmed colorectal cancer with liver metastases;
3. Immunohistochemistry (IHC) assessment shows GCC expression in tumor lesions of ≥1+ in more than 50% of the region (evaluation is performed by randomly selecting at least five tumor areas, and at least five unstained slides must be provided for assessment);
4. At least one measurable lesion;
5. Patients with colorectal cancer who have no standard treatment available, have progressed after third-line therapy, or are intolerant to third-line treatment;
6. Expected survival time of ≥90 days;
7. Normal function of major organs, as defined by the following criteria:

   1. Absolute neutrophil count (ANC) ≥1.5 x 10\^9/L;
   2. Platelets ≥80 x 10\^9/L;
   3. Hemoglobin ≥9 g/dL;
   4. Liver function:

      Total bilirubin ≤1.5 times the upper limit of normal (ULN), with subjects with Gilbert's syndrome having bilirubin ≤2.0 times ULN; Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) ≤1.5 times ULN (≤3.0 times ULN if liver metastases are present);
   5. International normalized ratio (INR) \<1.3; if the subject is receiving anticoagulant therapy, INR \<3 is required;
   6. Serum creatinine ≤1.5 mg/dL (132.6 µmol/L) or estimated glomerular filtration rate (eGFR) ≥50 mL/min/1.73 m²;
   7. Left ventricular ejection fraction (LVEF) \>50%;
8. No bleeding disorders or coagulation dysfunction;
9. Women of childbearing potential must undergo a pregnancy test (serum or urine) within 7 days prior to enrollment, with a negative result, and must be willing to use appropriate contraception during the trial and for 8 weeks after the last CAR-T administration (women who have undergone sterilization or have been postmenopausal for at least 2 years are considered non-childbearing);
10. Subjects must voluntarily participate in the study, sign the informed consent form, have good compliance, and be willing to cooperate with follow-up.

Exclusion Criteria:

1. Pregnant or breastfeeding women;
2. Receipt of chemotherapy, targeted therapy, monoclonal antibody therapy, or traditional Chinese anti-tumor medicine within 14 days prior to cell collection for enrollment;
3. Participation in another clinical drug trial within 4 weeks before the start of the study;
4. Presence of any of the following cardiovascular diseases or risk factors:

   1. Myocardial infarction, unstable angina, acute or persistent myocardial ischemia, Grade 3 or 4 heart failure (according to NYHA classification), severe symptomatic or poorly controlled arrhythmia, cerebrovascular accident, transient ischemic attack, or other serious cardiovascular diseases within 6 months prior to enrollment;
   2. History of myocarditis, primary cardiomyopathy, or specific cardiomyopathy;
   3. Disseminated intravascular coagulation (DIC), peripheral arterial thrombosis, pulmonary embolism, or other severe thromboembolic events within 3 months prior to enrollment;
   4. Aortic aneurysm, aortic dissection, or other life-threatening vascular diseases requiring surgery within 6 months prior to enrollment;
   5. QTcF interval \>480 ms;
   6. Left ventricular ejection fraction (LVEF) \<50% as shown by echocardiography (ECHO);
5. Long-standing unhealed wounds or fractures;
6. Presence of bleeding disorders or coagulation dysfunction;
7. History of substance abuse involving psychiatric medications, or a history of mental illness that cannot be controlled;
8. Uncontrolled or active fungal, bacterial, viral, or other infections;
9. Previous anti-tumor treatment toxicity that has not recovered to ≤ Grade 1 or to the levels specified in the inclusion criteria;
10. Known HIV infection; active syphilis infection; or active hepatitis B (HBsAg-positive, and HBV-DNA ≥500 IU/mL or above the lower limit of detection, whichever is higher) or hepatitis C virus infection (HCV antibody-positive, and HCV-RNA above the lower limit of detection);
11. Uncontrolled pleural effusion, pericardial effusion, or ascites requiring repeated drainage despite appropriate intervention;
12. History of severe allergic reactions to the main therapeutic drugs in this study (including fludarabine, cyclophosphamide, mesna, tocilizumab, and anti-infective drugs used during preconditioning);
13. Patients with active autoimmune diseases requiring systemic treatment within the past two years (including but not limited to autoimmune hepatitis, uveitis, enteritis, hypophysitis, vasculitis, nephritis, hyperthyroidism, etc.); hormone replacement therapy, such as thyroid hormone, insulin, or physiological corticosteroid replacement therapy for adrenal or pituitary insufficiency, is not considered systemic treatment;
14. Female subjects unwilling to use contraception from the time of signing the consent form until 6 months after CAR-T cell infusion;
15. Patients with meningeal, brainstem, spinal cord metastases, and/or compression, or active or untreated central nervous system (CNS) metastases;
16. History of interstitial lung disease (ILD) or non-infectious pneumonia requiring corticosteroid treatment;
17. Clinically severe lung damage caused by pulmonary complications, including but not limited to any underlying lung disease (e.g., pulmonary embolism within 3 months prior to enrollment, severe asthma, severe chronic obstructive pulmonary disease) or any autoimmune, connective tissue, or inflammatory disease affecting the lungs (e.g., rheumatoid arthritis, sarcoidosis, etc.), or previous total lung resection;
18. Any condition that the investigator believes could interfere with drug evaluation, compromise the subject's safety, or affect the study results, or any condition that the investigator deems unsuitable for participation in this study.