DNA Immunization in the Female Genital Tract Using A Controlled Delivery Matrix

描述（由申请人提供）：每年估计有239，000名妇女死于宫颈癌。超过99%的这些死亡是由生殖器感染人乳头瘤病毒（HPV）造成的。因此，迫切需要开发一种治疗性疫苗来解决由这种疾病引起的癌性病变。该提案的主要目标是开发一种解决HPV引起的癌前和癌性宫颈病变的设备。这项工作的一个目标是制造DNA疫苗输送装置局部施用在女性生殖道。此外，将进行体外和体内临床前研究，以评估该器械在接种有效疫苗方面的有效性。为此，已经提出了以下具体目标：（1）通过DNA的受控释放表征阴道内接种的分子和免疫学事件;（2）研究剂量、持续时间和来自DNA递送装置的转染剂对免疫学事件的影响;和（3）研究通过阴道内DNA递送的肿瘤消退的功效。将研究一种以受控方式释放DNA的装置。具体地，将质粒DNA掺入由以圆盘形状制造的生物相容性材料组成的基质中。该质粒DNA将编码HPV的致癌基因，其将充当治疗性疫苗以刺激免疫相关应答。临床前实验将在模型小鼠中进行，其中该装置将被插入阴道。将通过实时PCR评价生殖道中DNA和mRNA表达的持续性，以评估器械的释放。此外，还将评估对治疗的体液和细胞免疫应答。一旦对这些事件进行了表征，将进一步修改该装置，以探索剂量、释放持续时间和转染剂的添加对免疫应答的持续时间和幅度的影响。最后，将探索激发在幅度和持续时间上最大的反应的装置，用于解决小鼠模型中的HPV特异性肿瘤。在这些研究结束时，将阐明对阴道内DNA免疫的更好理解，并将开发解决HPV特异性病变的装置。解决宫颈病变的非侵入性装置通过提供不受熟练人员管理限制的治疗而对全世界妇女的健康具有直接益处。此外，这种疫苗接种策略可以用于开发其他性传播疾病的新疗法。

DESCRIPTION (provided by applicant): Each year an estimated 239,000 women die of cervical cancer. Over 99% of these deaths result from genital infection with human papillomavirus (HPV). As a result there is a pressing need to develop a therapeutic vaccine to resolve cancerous lesions caused by this disease. The broad goal of this proposal is to develop a device which will resolve pre-cancerous and cancerous cervical lesions caused by HPV. One objective of this work is to fabricate DNA vaccine delivery devices to be administered topically in the female genital tract. Furthermore, in vitro and in vivo preclinical studies will be performed to assess the efficacy of this device in administering an effective vaccine. To this end the following specific aims have been proposed: (1) characterize molecular and immunological events of intravaginal vaccination by controlled release of DNA; (2) investigate the influence of dosage, duration, and transfection agents from DNA delivery devices on immunological events; and (3) investigate the efficacy of tumor resolution by intravaginal DNA delivery. A device will be investigated which releases DNA in a controlled fashion. Specifically, plasmid DNA will be incorporated into a matrix comprised of a biocompatible material fabricated in the shape of a disk. This plasmid DNA will encode an oncogene of HPV which will act as a therapeutic vaccine to stimulate immune relevant responses. Pre-clinical experiments will be implemented in a model mouse, where the device will be inserted in the vagina. Persistence of DNA and mRNA expression in the genital tract will be evaluated by real-time PCR to assess the release from the device. Additionally, both humoral and cellular immune responses to the therapy will be assessed. Once these events have been characterized the device will be further modified to explore the influence of dose, duration of release, and addition of transfection agents on duration and magnitude of immune response. Finally, the device that elicits a response that is largest in magnitude and duration will be explored for resolving HPV specific tumor in a mouse model. At the conclusion of these studies a better understanding of intravaginal DNA immunization will have been elucidated and processes will be made for developing a device to resolve HPV specific lesions. A non-invasive device to resolve cervical lesions has immediate benefit to women's health worldwide by providing a therapy that is not limited by skilled personal to administer. Additionally, this vaccination strategy could be applied to develop new therapies for other sexually transmitted diseases.