CELL INTELLIGENCE · 肿瘤细胞治疗研究

Inclusion Criteria:

* Age: ≤ 40 years at the time of recurrence or progression with any type of sarcoma that has recurred or not responded to standard therapy and is deemed incurable by standard therapy.
* Positive NKG2DL expression in sarcoma samples. Ideally, they should have centralized histological verification of NKG2DL expression in sarcoma samples (positive expression is defined as at least 2+ expression (0-4+ scale) in \>50 percent of the tumor cells using anti-MICA and or anti-ULBP2). Patients will undergo biopsy following enrollment to obtain tissue to assess NKG2DL expression, with the following restrictions:

  * If the patient doesn´t have adequate accessible tumor for biopsy (at least 1 cm diameter).
  * Procedures employed to acquire biopsies for tumor lysates will be limited to percutaneous needle or core biopsies, thoracoscopic excision or open biopsies of readily accessible lesions. Pulmonary lesions may be biopsied but extensive surgery such as thoracotomy or laparotomy should not be employed.
  * Patients who require biopsy should not be enrolled if in the opinion of the principal investigator (PI), the tumor site places the patient at substantial related risk from the biopsy procedure.

In patients that fulfill any of these restrictions, when adequate archived tissue is available, this may be utilized to assess NKG2DL expression.

* Patient must have either measurable or evaluable tumor.
* The tumor must be accessible for intralesional administration of CAR T cells (only in ARM B).
* Life expectancy of at least 10 weeks in opinion of the principal investigator (PI).
* Lansky (age \<16 years) or Karnofsky (age \>=16 years) score of 50 or greater.
* Patients must have recovered from the acute toxic effects of all prior anticancer therapy (including chemotherapy and radiotherapy).
* Adequate bone marrow function defined by an absolute neutrophil count (ANC) of \>/= 1.000/μL, platelet count of \>/= 30.000/μL and hemoglobin of \>/= 9.0 g/dl, and absence of a regular red blood cell and platelet transfusion requirement.
* Patients should have a normal hepatic function with a total bilirubin \<2 times the upper limit of normal and serum glutamic oxaloacetic transaminase (SGOT) or serum glutamic pyruvic transaminase (SGPT) \< 2 times the upper limit of normal, and adequate renal function as defined by a serum creatinine ≤ 1.5 upper limit of normal.
* Patient or patient's legal representative, parent(s), or guardian able to provide written informed consent.
* Sexually active patients must be willing to utilize one of the more effective birth control methods for 6 months after the infusion. Male partner should use a condom.

Women of child-bearing potential, defined as all women physiologically capable of becoming pregnant, unless they are using highly effective methods of contraception while taking study treatment and for at least 6 months after the NKG2D-CAR T infusion and until CAR-T cells are no longer present by qPCR on two consecutive tests. Highly effective contraception methods include, as defined by the CTFG recommendations (available at h t t p s : / / w w w . h m a . e u / f i l e a d m i n / d a t e i e n / H u m a n \_ M e d i c i n e s / 0 1 -About\_HMA/Working\_Groups/CTFG/2014\_09\_HMA\_CTFG\_Contraception.pdf):

* Combined (estrogen and progestogen containing) hormonal contraception associated with inhibition of ovulation (oral, intravaginal, transdermal)
* Progestogen-only hormonal contraception associated with inhibition of ovulation (oral, injectable, implantable)
* Intrauterine device (IUD)
* Intrauterine hormone-releasing system
* Bilateral tubal occlusion
* Vasectomised partner (provided that partner is the sole sexual partner of the trial participant and that the vasectomised partner has received medical assessment of the surgical success)
* Sexual abstinence (only if defined as refraining from heterosexual intercourse during the entire period of risk associated with the study treatments. The reliability of sexual abstinence needs to be evaluated in relation to the duration of the clinical trial and the preferred and usual lifestyle of the subject).

Sexually active males should use a condom during intercourse while taking study treatment and for at least 6 months after the infusion and until CAR-T cell are no longer present by qPCR on two consecutive tests.

Exclusion Criteria:

* Enrolled in another treatment protocol.
* Evidence of untreated and active infection or clinically significant systemic illness:

  * Cardiac disorder defined as LVFE \< 45% determined by ECHO.
  * Human Immunodeficiency Virus (HIV) positive test.
  * Presence of active or prior CMV, EBV, hepatitis B or C as indicated by serology.
  * Any significant pulmonary, hepatic or other organ dysfunction.
* Chronic corticosteroid dependence (except replacement therapy).
* Evidence of any toxicity grade ≥ 4 (according to Common Terminology Criteria for Adverse Events (CTCAE) Version 5.0).
* Pregnant or lactating women.
* Medical history of epilepsy.
* Any other condition that, in the opinion if the PI, may interfere with the efficacy and/or safety evaluation of the trial.

主要终点：Safety: Dose-limiting toxicity (DLT) of NKG2D-CAR memory T cells · 1. Any grade 3 or higher toxicity with an attribution of definitely or probably related to the infusion of the NKG2D CAR-T cells with the exception of Grade 3 fever and immediate hypersensitivity reactions occurring within 2hours of cell infusion that are reversible to a Grade 2 or less within 24 hours of cell administration with standard therapy; 2. Any lower grade toxicity that increases to a grade 3 or higher as a direct result of the NKG2D CAR-T cell infusion. · During the study treatment, until 28 days after the last study iv treatment administration；Safety: Maximum Tolerated Dose (MTD) of NKG2D-CAR memory T cells · The highest dose level if no Dose-limiting toxicitys are observed · During the study treatment, until 28 days after the last study iv treatment administration；Response rate · This outcome will be evaluated by Immune-Related Response Criteria (iRECIST) v1.1. The efficacy will be measured by objective response rate (ORR) at D60 in both arms, which includes Complete Response (CR) and Partial Response (PR) based on Immune-Related Response Criteria iRECIST v1.1. · At day 60 after the treatment
次要终点：Rate of NKG2D-CAR T cells persistence in peripheral blood；Rate of NKG2D-CAR T cells persistence in the tumor and metastasis site；Rate of NKG2DL positive expression on primary sarcoma samples；Cytokine determination in the serum of patients；Analysis of patient peripheral blood immune cell subpopulations；Identify the DNA methylation profile of NKG2DL (MICA, MICB AND ULBPS 1-3) in primary sarcoma samples and DNA methylation profile of NKG2D-T cells before and after infusion.；Evaluate the presence of soluble NKG2DL and ANTI-MICA antibodies in the serum of patients under therapy

• NKG2D-CAR memory T cells infusion · EXPERIMENTAL · If the primary tumor and/or metastases are not accessible, the patient will be included in Arm A.This group will receive an intravenous infusion of NKG2D-CAR memory T cells.
• Systemic and locally transduced NKG2D-CAR memory T cells infusion · EXPERIMENTAL · If the primary tumor and/or metastases are accessible, the patient will be included in Arm B. This group will receive an intravenous infusion of NKG2D-CAR memory T cells and an intratumoral dose of NKG2D-CAR memory T cells.

Phase I, open label, prospective, single-center, non-randomized, dose escalation clinical trial aiming to determine the dose-limiting toxicity (DLT) and maximum tolerated dose (MTD) of systemic transduced donor-derived NKG2D-CAR memory T cell infusions (Arm A), and of dual treatment, with both systemic and locally transduced donor-derived NKG2D-CAR memory T cell infusions (Arm B).