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

描述(申请人提供)：每年估计有239,000名妇女死于宫颈癌。其中99%以上的死亡是由生殖器感染人乳头瘤病毒(HPV)引起的。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症损害。这项提案的主要目标是开发一种设备，用于解决HPV引起的癌前病变和癌前病变。这项工作的一个目标是制造DNA疫苗递送装置，用于女性生殖道的局部注射。此外，将进行体外和体内的临床前研究，以评估该设备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)通过控制释放DNA来表征阴道内接种的分子和免疫学事件；(2)研究DNA递送装置中的剂量、持续时间和转染剂对免疫事件的影响；以及(3)通过经阴道递送DNA来研究肿瘤消退的效果。将对一种以受控方式释放DNA的设备进行调查。具体地说，质粒DNA将被加入到由制造成圆盘形状的生物兼容材料组成的基质中。该质粒DNA将编码HPV的癌基因，作为治疗性疫苗刺激免疫相关反应。临床前实验将在模型小鼠身上进行，该设备将被插入阴道。实时定量聚合酶链式反应将评估DNA和信使核糖核酸在生殖道的持续表达，以评估该设备的释放情况。此外，还将评估对该疗法的体液和细胞免疫反应。一旦这些事件被表征，该装置将被进一步改进，以探索剂量、释放持续时间和转染剂添加对免疫反应持续时间和大小的影响。最后，将探索在小鼠模型中用于解决HPV特异性肿瘤的装置，该装置在幅度和持续时间上引起最大的反应。在这些研究的结论中，将对阴道内DNA免疫有更好的了解，并将开发一种设备来解决HPV特异性病变。一种解决宫颈病变的非侵入性设备提供了一种不受熟练个人管理限制的治疗方法，对全球妇女的健康具有直接的好处。此外，这一疫苗接种策略可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.