CELL INTELLIGENCE · 肿瘤细胞治疗研究

Inclusion Criteria:

* Age: 18 to 70 years old (inclusive); gender unrestricted.

Diagnosis of advanced Hepatocellular Carcinoma (HCC), meeting the following requirements:

* Pathologically Confirmed: Diagnosis of HCC confirmed by histopathology. Staging: Classified as China Liver Cancer (CNLC) stage IIb-IIIb, having undergone treatments recommended by the "Primary Liver Cancer Diagnosis and Treatment Guidelines (2024 Edition)" with disease progression and either no further recommended treatments available or intolerance to the recommended treatment options.
* Measurable Lesion: At least one measurable lesion as defined by RECIST v1.1 criteria.
* Tumor Sample Availability: Availability of tumor tissue samples or samples obtained by tumor biopsy for GPC3 expression quantification and other related analyses.
* GPC3 Positivity: Confirmed positive GPC3 expression by immunohistochemistry (IHC), where positivity is defined as a quantified immunohistochemical score of "+" or above.
* ECOG Performance Status: Eastern Cooperative Oncology Group (ECOG) score of 0-1.
* Life Expectancy: Expected survival time of ≥ 3 months.
* Cirrhosis Status: Child-Pugh class A or B for liver cirrhosis.
* Organ Function: Must meet the following organ function requirements:

Hematology:

Absolute neutrophil count (ANC) ≥ 1.5 × 10\^9/L (no granulocyte colony-stimulating factor support within 7 days prior to testing).

Absolute lymphocyte count (ALC) ≥ 0.5 × 10\^9/L; hemoglobin (HGB) ≥ 80 g/L (no red blood cell transfusion within 7 days prior to testing).

Platelet count (PLT) ≥ 75 × 10\^9/L (no transfusion support within 7 days prior to testing).

Liver Function:

Aspartate aminotransferase (AST ) and alanine aminotransferase (ALT ) ≤ 3.0 × upper limit of normal (ULN).

Total bilirubin (TBIL) ≤ 2.0 × ULN (≤ 3.0 × ULN for patients with Gilbert's syndrome and direct bilirubin ≤ 1.5 × ULN).

Coagulation Function:

International normalized ratio (INR) ≤ 1.5 × ULN. Activated partial thromboplastin time (APTT) ≤ 1.5 × ULN (except for patients receiving therapeutic anticoagulants).

Renal Function: Serum creatinine (Cr) ≤ 1.5 × ULN or creatinine clearance ≥ 60 mL/min.

Cardiac Function: Left ventricular ejection fraction (LVEF) ≥ 50% (confirmed by echocardiography).

Pulmonary Function: Pulse oxygen saturation (SpO2) \> 93% at rest without supplemental oxygen.

* Contraception: Women of childbearing potential must have a negative pregnancy test, and both male and female participants with reproductive potential must agree to use effective contraception throughout the screening and study period until one year after the last cellular infusion.
* Informed Consent: Willingness to provide voluntary written informed consent and compliance with the study protocol.

Exclusion Criteria:

* Pregnant or breastfeeding women.
* Positive HCV RNA quantification, positive human immunodeficiency virus (HIV) antibodies, or active syphilis infection.
* Chronic HBV infection with serum HBV-DNA levels ≥ 500 IU/mL.
* Unresolved non-hematologic toxicities (excluding alopecia and peripheral sensory neuropathy) from prior treatments (surgery, chemotherapy, radiotherapy, targeted therapy, immunotherapy) that have not improved to ≤ Grade 1 according to CTCAE.
* History of allogeneic tissue/organ transplantation (including bone marrow, stem cell, liver, or kidney transplants), except those that do not require immunosuppressive therapy (e.g., corneal or hair transplants).
* Prior treatment targeting GPC3.
* Receipt of anti-tumor treatment for liver cancer or any other medical intervention that could impair major organ function within four weeks before signing informed consent.
* Known central nervous system metastasis.
* Presence of clinically significant systemic disease (e.g., severe active - - infections, significant heart, lung, liver, kidney, or neurological dysfunction) that, in the investigator's opinion, may impair the patient's ability to tolerate the study treatment or increase the risk of complications. Including but not limited to:

  1. Uncontrolled severe active infection.
  2. Symptomatic congestive heart failure (NYHA Class II-IV).
  3. Clinically significant severe aortic valve stenosis or symptomatic mitral valve stenosis.
  4. QTc \> 450 msec on ECG, or QTc \> 480 msec in patients with bundle branch block.
  5. Uncontrolled clinically significant arrhythmias within six months before signing informed consent.
  6. Acute coronary syndrome (e.g., unstable angina or myocardial infarction) within six months before signing informed consent.
  7. Hypertension not controlled by medication (systolic BP ≥ 160 mmHg and/or diastolic BP ≥ 100 mmHg).
  8. Cerebrovascular accidents, including transient ischemic attack (TIA), cerebral infarction, cerebral hemorrhage, or subarachnoid hemorrhage within six months before signing informed consent.
  9. Active, chronic, or recurrent severe autoimmune disease (within one year before signing informed consent), or liver cirrhosis/liver cancer caused by autoimmune hepatitis.
  10. Any form of primary or secondary immunodeficiency, such as severe combined immunodeficiency (SCID).
  11. Risk of organ perforation or hemorrhage, as determined by the investigator.
* History of severe systemic hypersensitivity to study drugs/components \[e.g., fludarabine, cyclophosphamide, dimethyl sulfoxide (DMSO), low molecular weight dextran, human serum albumin (HSA)\].
* Receipt of live attenuated vaccine within four weeks before signing informed consent.
* Participation in another clinical trial within four weeks before signing informed consent.
* History of another malignancy within the past five years, excluding adequately treated non-melanoma skin cancer or carcinoma in situ (e.g., breast, stomach, colon, bladder, cervix, or melanoma).
* History of neuropsychiatric disorders diagnosed by ICD-11 criteria, or any neuropsychiatric disorder deemed by the investigator to warrant exclusion, including but not limited to epilepsy, schizophrenia, dementia, or addiction to drugs/alcohol.
* Any other condition that, in the investigator's opinion, makes the patient unsuitable for this clinical trial.

主要终点：Incidence of treatment-related adverse events · Treatment-related adverse events are defined as any medical events occurring since the initiation of GPC3-targeted CAR T cell therapy. CRS or CRES will be graded based on the American Society for Transplantation and Cellular Therapy (ASTCT) criteria, and other adverse events will be graded according to CTCAE v5.0. · Up to 12 months since the initiation of GPC3-targeted CAR T cell therapy.；Incidence of dose-limiting toxicities (DLTs) · Dose-limiting toxicities are defined as GPC3-targeted CAR T cell therapy-related adverse events within the first 28 days that meet the following criteria: grade 3 or higher CRS or CRES, and any other grade 4 adverse events. · Up to 28 days from the initiation of GPC3-targeted CAR T cell therapy.
次要终点：Number and copy number of GPC3-targeted CAR T cells；Objective response rate (ORR)；Progression Free Survival (PFS)；Time to response (TTR)；Duration of response (DOR)；Overall Survival (OS)

• GPC3 CAR-T cell therapy · EXPERIMENTAL · Enrolled patients will receive a single infusion of GPC3 CAR-T cells at a starting dose of 1×10\^6 cells/kg.

In this single-center, single-arm, prospective, open-label Phase 1/2 study, the safety and efficacy of autologous GPC3-targeted chimeric antigen receptor (CAR) T-cell therapy will be evaluated in patients with GPC3-positive advanced hepatocellular carcinoma. Phase 1 will involve the enrollment of six eligible patients to receive hepatic arterial infusion of GPC3-CAR T cells at a fixed dose of 1×10\^6 cells/kg, with or without a standard lymphodepleting conditioning regimen (fludarabine and cyclophosphamide). Based on the results, it will be assessed whether the FC lymphodepletion regimen is necessary. Subsequently an additional six patients will be enrolled in a "3+3" dose-escalation design to adjust the dose of GPC3-CAR T cells to achieve optimal safety and efficacy. The recommended Phase 2 dose (RP2D) will then be established. Phase 2 will involve the enrollment of 10-20 additional eligible patients to receive GPC3-CAR T cell therapy at the RP2D.