Mahvi D M, Sondel P M, Yang N S, Albertini M R, Schiller J H, Hank J, Heiner J, Gan J, Swain W, Logrono R
Department of Surgery, H4/726 Clinical Science Center, Madison, Wisconsin 53792, USA.
Hum Gene Ther. 1997 May 1;8(7):875-91. doi: 10.1089/hum.1997.8.7-875.
The objective of this Phase I study is to assess the acute and long-term toxicities of intradermal vaccination of cancer patients with lethally-irradiated tumor cells that have been transfected by particle-mediated gene transfer (PMGT) with gold particles coated with human granulocyte-macrophage colony stimulating factor (GM-CSF) DNA in a plasmid expression vector. The GM-CSF DNA-coated gold particles are delivered to tumor cells using helium pressure with a hand held gene delivery device. Preclinical studies have demonstrated that vaccination of mice with irradiated, GM-CSF-transfected melanoma cells provided protection from subsequent challenges with non-irradiated, non-transfected tumor cells. Ongoing human tumor immunotherapy studies use patients' melanoma or renal carcinoma cells transfected with a retroviral vector containing GM-CSF cDNA as a vaccine to elicit anti-tumor immune responses. PMGT transfection, unlike retroviral transfection, does not require tumor cells to proliferate in vitro to undergo gene transfer. Instead, tumor tissue can be dissociated into small tissue clumps or cell aggregates and then immediately transfected using the gene gun. PMGT physically inserts the DNA without the need for cell surface interaction with viral components or exposure of the patient to viral antigens. As described in this protocol, fresh human sarcoma and melanoma specimens can be transfected with the GM-CSF DNA-coated gold particles with subsequent production of biologically active GM-CSF protein. In this study tumor tissue will be obtained from patients with melanoma or sarcoma. Tumor tissue will be dissociated, irradiated, and transfected with GM-CSF DNA by PMGT. In this ascending dosage study, two dose levels of GM-CSF DNA will be studied in 2 groups of 6 patients each. Patients will receive two intradermal injections of the irradiated, transfected tumor in a single extremity. On days 3 and 14 post-vaccination, patients will undergo surgical excision of the vaccination sites to assess GM-CSF production and infiltration of immune effector cells. On Day 25, patients will undergo DTH testing with intradermal injection in their opposite extremity of 5 x 10(6) irradiated non-transfected autologous tumor cells cryopreserved at the time of vaccine preparation. This injection site will be assessed on day 28 post-vaccination and surgical excision of the DTH testing site will be performed on day 28 if a positive reaction is noted. The patients will be observed for local and systemic toxicity on days 2, 3, 5, 8, 14, 25, and 28 after the vaccination. Restaging of the patients' disease and long term toxicity evaluation will be performed at 3, 6, and 12 months and then yearly.
本I期研究的目的是评估癌症患者皮内接种经粒子介导基因转移(PMGT)转染的致死性照射肿瘤细胞的急性和长期毒性,这些肿瘤细胞用涂有人类粒细胞巨噬细胞集落刺激因子(GM-CSF)DNA的金颗粒在质粒表达载体中进行转染。使用手持式基因递送装置通过氦气压力将涂有GM-CSF DNA的金颗粒递送至肿瘤细胞。临床前研究表明,用照射过的、GM-CSF转染的黑色素瘤细胞对小鼠进行疫苗接种可使其免受随后未照射、未转染的肿瘤细胞的攻击。正在进行的人类肿瘤免疫治疗研究使用用含有GM-CSF cDNA的逆转录病毒载体转染的患者黑色素瘤或肾癌细胞作为疫苗来引发抗肿瘤免疫反应。与逆转录病毒转染不同,PMGT转染不需要肿瘤细胞在体外增殖即可进行基因转移。相反,肿瘤组织可以解离成小的组织块或细胞聚集体,然后立即使用基因枪进行转染。PMGT通过物理方式插入DNA,无需细胞表面与病毒成分相互作用或使患者暴露于病毒抗原。如本方案所述,新鲜的人类肉瘤和黑色素瘤标本可用涂有GM-CSF DNA的金颗粒进行转染,随后产生具有生物活性的GM-CSF蛋白。在本研究中,肿瘤组织将取自黑色素瘤或肉瘤患者。肿瘤组织将被解离、照射,并通过PMGT用GM-CSF DNA进行转染。在这项剂量递增研究中,将在两组各6名患者中研究两个剂量水平的GM-CSF DNA。患者将在单个肢体接受两次皮内注射经照射、转染的肿瘤。在接种疫苗后的第3天和第14天,患者将接受接种部位的手术切除,以评估GM-CSF的产生和免疫效应细胞的浸润。在第25天,患者将在对侧肢体进行皮内注射5×10⁶个在制备疫苗时冷冻保存的经照射、未转染的自体肿瘤细胞,进行迟发型超敏反应(DTH)测试。该注射部位将在接种疫苗后的第28天进行评估,如果出现阳性反应,将在第28天对DTH测试部位进行手术切除。在接种疫苗后的第2、3、5、8、14、25和28天观察患者的局部和全身毒性。在3、6和12个月,然后每年对患者的疾病进行重新分期并进行长期毒性评估。
Oncoimmunology. 2019-1-11
Cytotechnology. 2001-7