Laser treatment to enhance therapeutic HPV DNA vaccine potency

激光治疗可增强 HPV DNA 疫苗的治疗效力

• 批准号: 8075018
• 负责人: TZYY-CHOOU WU
• 金额: $ 24.88万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8075018
• 关键词: Antigen Presentation Pathway; Antigen-Presenting Cells; Antigens; Area; Biological Assay; Bone Marrow; C57BL/6 Mouse; CD8B1 gene; CTL assay; Cancer Etiology; Cells; Cellular Immunity; Cervical; Cessation of life; Clinical; DNA; DNA Vaccines; Dendritic Cells; Dendritic cell activation; Devices; Dose; Foundations; Future; Gene Delivery; Histocompatibility Antigens Class II; Human Papilloma Virus Vaccine; Human Papillomavirus; Human papilloma virus infection; Human papillomavirus 16; Immune; Immune response; Immunotherapy; Injection of therapeutic agent; Langerhans cell; Lasers; Lead; Lesion; Link; Luciferases; Lung; MHC Class I Genes; MHC Class II Genes; Maintenance; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Mediating; Methods; Modeling; Mus; Muscle; Needles; Neoplasm Metastasis; Oncogenic; Play; Proteins; Regimen; Role; Skin; Staining method; Stains; System; T cell response; T-Lymphocyte; Techniques; Technology; Testing; Therapeutic; Transfection; Translating; Translations; Tumor Cell Line; Tumor Immunity; Vaccinated; Vaccination; Woman; calreticulin; cell mediated immune response; cytokine; enzyme linked immunospot assay; gene gun; head-to-head comparison; immunogenicity; improved; in vivo; innovation; interest; intradermal injection; keratinocyte; killer T cell; lymph nodes; metaplastic cell transformation; novel strategies; prevent; public health relevance; research study; subcutaneous; tumor; tumor growth; vaccination strategy

DESCRIPTION (provided by applicant): Human papillomavirus E7 oncogenic protein is consistently expressed in HPV-expressing cervical cancers and is important in the induction and maintenance of cellular transformation. Thus, immunotherapy targeting E7 may provide an opportunity to treat HPV-associated cervical malignancies. We have previously shown that intradermal gene gun delivery of DNA vaccines encoding calreticulin (CRT) linked to E7 (CRT/E7) can significantly enhance E7-specific CD8+ T cell immune responses and antitumor effects against E7-expressing tumors. One major limitation of this DNA vaccination strategy is the limited transfection efficiency in vivo, which may limit the potency of DNA vaccines. Recent studies have described a novel approach to increase the transfection efficiency of the DNA vaccines in vivo using the femtosecond laser beam treatment. Thus, in the current proposal, we hypothesize that intradermal administration of the CRT/E7 DNA vaccine via needle injection or gene gun followed by femtosecond laser beam treatment would significantly enhance the transfection efficiency of the DNA vaccine, resulting in enhanced E7-specific T cell-mediated immune responses and antitumor effects in vivo. Specifically, we propose to: Specific Aim 1: Characterize the E7- specific CD8+ T cell immune responses and antitumor effects against an E7-expressing tumor using intradermal needle injection of the CRT/E7 DNA vaccine with or without laser treatment. Specific Aim 2: Characterize the E7-specific CD8+ T cell immune response and antitumor effects against an E7-expressing tumor using gene gun administration of the CRT/E7 DNA vaccine with or without laser treatment. Specific Aim 3: Perform a head-to-head comparison of intradermal needle injection of CRT/E7 DNA vaccine followed by laser treatment with gene gun administration of CRT/E7 DNA vaccine followed by laser treatment for their ability to generate E7-specific CD8+ T cell immune responses and antitumor effects against E7-expressing tumors. Specific Aim 4: Determine the mechanism of how laser treatment leads to enhancement in antigen- specific CD8+ T cell immune responses and antitumor effects in treated mice. The successful implementation of this study will represent an innovative approach for enhancing DNA vaccine potency. In addition, this study will potentially serve as an important foundation for future clinical translation. PUBLIC HEALTH RELEVANCE: Cervical cancer is the second leading cause of cancer deaths in women worldwide and human papillomavirus (HPV) has been identified as a necessary cause of cervical cancer. Our project aims to develop an innovative therapeutic HPV DNA vaccination strategy employing laser technology to improve DNA vaccine potency. Successful implementation of this study will represent an innovative approach for enhancing the potency of DNA vaccines targeting HPV for the treatment of cervical cancer.

描述（由申请人提供）：人乳头瘤病毒E7致癌蛋白始终在表达HPV的宫颈癌中表达，并且在细胞转化的诱导和维持中很重要。因此，靶向E7的免疫疗法可以提供治疗与HPV相关的宫颈恶性肿瘤的机会。我们先前已经表明，与E7（CRT/E7）相关的编码钙网蛋白（CRT）的DNA疫苗的射传射击可以显着增强E7特异性CD8+ T细胞免疫反应和针对E7-表达肿瘤的抗肿瘤作用。该DNA疫苗接种策略的主要局限性是体内的转染效率有限，这可能会限制DNA疫苗的效力。最近的研究描述了一种使用飞秒激光束处理在体内提高DNA疫苗转染效率的新方法。因此，在当前的提案中，我们假设通过针注射或基因枪对CRT/E7 DNA疫苗的施用，然后进行巨秒激光束治疗将显着提高DNA疫苗的转染效率，从而增强E7特异性T细胞介导的免疫反应和抗毒剂的E7特异性T细胞介导的效果。具体而言，我们提出：特定目标1：通过使用激光治疗或无激光治疗的CRT/E7 DNA疫苗对E7特异性CD8+ T细胞免疫反应和抗肿瘤作用，对表达E7表达肿瘤的抗肿瘤作用。具体目标2：使用有或没有激光治疗的CRT/E7 DNA疫苗的基因枪施用E7特异性CD8+ T细胞免疫反应和抗肿瘤对表达E7表达肿瘤的作用。具体目标3：对CRT/E7 DNA疫苗的皮内针注射的头对头进行比较，然后用CRT/E7 DNA疫苗的基因枪对激光治疗进行激光治疗，然后对激光治疗进行激光处理，以产生E7特异性CD8+ T细胞免疫反应，并对E7表达E7-Excancepress Excancepress Excance tumers产生E7特异性CD8+ T细胞免疫反应。具体目标4：确定激光治疗如何导致抗原特异性CD8+ T细胞免疫反应和治疗小鼠抗肿瘤作用的机制。这项研究的成功实施将代表增强DNA疫苗效力的创新方法。此外，这项研究将有可能成为未来临床翻译的重要基础。 公共卫生相关性：宫颈癌是全球女性癌症死亡的第二大主要原因，而人类乳头瘤病毒（HPV）已被确定为宫颈癌的必要原因。我们的项目旨在开发一种使用激光技术来提高DNA疫苗效力的创新治疗HPV DNA疫苗接种策略。这项研究的成功实施将代表一种创新的方法，用于增强针对HPV治疗宫颈癌的DNA疫苗的效力。