Phase I study of intravesical vaccinia virus as a vector for gene therapy of bladder cancer
Phase I study of intravesical vaccinia virus as a vector for gene therapy of bladder cancer
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DOI：
10.1016/s0022-5347(05)65755-2
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发表时间：
2001-10-01
影响因子：
6.6
通讯作者：
Lattime, EC
中科院分区：
文献类型：
--
作者：
Gomella, LG;Mastrangelo, MJ;Lattime, EC
目的：痘苗病毒是一种DNA痘病毒，以前被用作根除天花的疫苗。该病毒感染效率高，无需染色体整合即可在细胞质内复制，并可运输大量重组DNA而不丧失感染性。因此，它是一种理想的体内基因传递载体。大量的痘苗疫苗已经注射到皮肤、皮下和周围淋巴结转移的黑色素瘤中，没有明显的副作用，并具有有效的肿瘤细胞感染和重组基因转染。为了确定膀胱内注射牛痘疫苗是否能有效地感染膀胱粘膜和肿瘤，并接受可接受的毒性，我们在根治性膀胱切除术前对肌肉浸润性移行细胞癌患者进行了I期膀胱内注射牛痘疫苗试验。材料和方法：在记录了免疫能力和复种后的主要反应后，患者接受了由疾病控制中心提供的3种递增剂量的水痘病毒膀胱内注射(惠氏-艾尔斯特实验室，费城，宾夕法尼亚州)。第三次注射后约24小时行膀胱切除术，并对组织进行显微镜检查。结果：共治疗4例。接受最高剂量(100×10(6)个斑块形成单位)的3名患者有显著的粘膜和粘膜下炎性细胞淋巴细胞、嗜酸性粒细胞和浆细胞渗入肿瘤和正常组织。树突状细胞在接触牛痘病毒后被招募到该部位。可见明显的粘膜水肿和血管扩张。肿瘤和正常的尿路上皮细胞显示有病毒感染的证据，包括增大的空泡细胞和胞浆内含物。除轻度排尿困难外，未见痘苗相关毒性的临床或实验室表现。在4例患者中，3例存活，4年后无病。结论：我们的研究表明，痘苗病毒可以安全地注射到膀胱内，通过淋巴细胞的募集和诱导活跃的局部炎症反应。据我们所知，这是第一份将活病毒直接送入人膀胱的报告。野生型牛痘疫苗在膀胱癌免疫治疗中的作用值得进一步研究。此外，这些数据支持探索重组牛痘疫苗作为膀胱内感染的潜在基因治疗载体，并将细胞因子或其他基因导入膀胱肿瘤细胞，这是我们团队开创的一种方法，最近在浅表性黑色素瘤患者中进行了研究。
Purpose: Vaccinia virus is a DNA poxvirus previously used as a vaccine to eradicate smallpox. The virus has a high efficiency of infection, replicates in the cytoplasm without chromosomal integration and can transport a large amount of recombinant DNA without losing infectivity. Therefore, it is an excellent choice as a vector for gene delivery in vivo. Large quantities of vaccinia have been injected into dermal, subcutaneous and peripheral lymph node melanoma metastases without significant side effects, and with efficient infection of the tumor cells and recombinant gene transfection. To determine if vaccinia, when given intravesically, can effectively infect bladder mucosa and tumor with acceptable toxicity, we performed a phase I trial of intravesical vaccinia in patients with muscle invasive transitional cell carcinoma before radical cystectomy.Materials and Methods: After documenting immune competence and demonstration of a major reaction after revaccination, patients received 3 increasing doses of intravesical Dryvax vaccinia virus (Wyeth-Ayerst Laboratories, Philadelphia, Pennsylvania) that was provided by the Centers for Disease Control. Approximately 24 hours after the third dose, cystectomy was performed and the tissue was examined microscopically.Results: There were 4 patients who were treated. The 3 patients who received the highest doses (100 x 10(6) plaque forming units) had significant mucosal and submucosal inflammatory infiltration by lymphocytes, eosinophils, and plasma cells into tumor and normal tissue. Dendritic cells were recruited to the site after exposure to the vaccinia. Significant mucosal edema and vascular ectasia were seen. Tumor and normal urothelial cells showed evidence of viral infection, including enlarged vacuolated cells with cytoplasmic inclusions. There were no clinical or laboratory manifestations of vaccinia related toxicity except mild dysuria. Of the 4 patients 3 survived and were free of disease at 4-year followup.Conclusions: Our study demonstrates that vaccinia virus can be administered safely into the bladder with recruitment of lymphocytes and induction of a brisk local inflammatory response. To our knowledge, this is the first report of direct delivery of live virus into the human bladder. The role of wild type vaccinia as immunotherapy for bladder cancer warrants further study. Furthermore, these data support the exploration of recombinant vaccinia as a putative gene therapy vector for intravesical infection and transfection of bladder tumor cells with cytokine or other genes, an approach that our group pioneered and most recently studied in patients with superficial melanoma.