Targeted-and Image-Based Adenovirus Cancer Therapeutic Vectors

描述(由申请人提供)：我们假设一种新的衣壳掺入方法将允许在体内全面监测癌症重定向载体，并允许评估我们新的载体工程策略，以实现对播散性肿瘤疾病的靶向治疗。通过研究腺病毒(Ad)载体的设计，我们预计从这一倡议中获得的新信息将证明我们高度新颖的成像方法的实用性，并验证重定向肿瘤转导的特异性。该应用程序的目标是开发一种结合了成像和靶向治疗的多功能Ad载体。为了实现这一目标，我们将结合三项独特的技术来推进播散性肿瘤疾病的治疗靶向：癌症特异性重定向、肝脏去靶向和非侵入性成像。我们通过将靶向单链抗体(ScFv)整合到病毒衣壳中，并结合用于肝脏去靶向和逃避先前存在的免疫的六邻体修饰，开发了能够细胞特异性靶向的Ad载体。我们创新的Ad靶向方法提供了一种新的方法来避免Ad和复杂的靶向配体(如scFv)之间的结构和生物合成不相容的问题，并可以使用包括重组抗体和生长因子在内的新型靶向配体来促进Ad靶向广泛的临床重要细胞群。本应用的目标是开发一种结合成像和靶向治疗的多功能Ad载体。为了实现这一目标，我们将结合三项独特的技术来推进播散性肿瘤疾病的治疗靶向：癌症特异性重定向、肝脏去靶向和非侵入性成像。我们通过将靶向单链抗体(ScFv)整合到病毒衣壳中，并结合用于肝脏去靶向和逃避先前存在的免疫的六邻体修饰，开发了能够细胞特异性靶向的Ad载体。我们的创新的Ad靶向方法提供了一种新的方法来避免Ad和复杂的靶向配体(如scFv)之间的结构和生物合成不相容的问题，并可以促进Ad靶向多种临床重要的细胞群。我们创新地使用了将金属硫蛋白整合到腺病毒Pix基因中的结构融合蛋白，提供了非侵入性的成像优势，即在给药后检测Ad载体的物理生物分布和扩散，这是使用报告基因不可能的。此外，能够在全身水平上非侵入性地观察Ad功能，使得有可能检测到肿瘤部位之外的病毒传播(S)，以监测临床安全性。通过在Ad载体设计中结合这些修改，我们预计从这一倡议中获得的新信息将展示我们高度新颖的成像方法的实用性，并验证靶向肿瘤转导的特异性。这项工作解决了晚期转移性癌症新疗法尚未得到满足的重大需求。所提出的成像方法的新颖设计不同于先前描述的载体成像模式，后者一直完全基于监测报告基因的表达。此外，我们的联合技术对基因治疗的不同领域产生了广泛的影响，在这些领域中，普遍需要实现细胞特异性转导和监测的能力。为了实现这些目标，我们组建了一支出色的研究团队，他们拥有腺病毒载体设计和构建、腺病毒重定向方法、使用Ad载体的临床前模型、非侵入性PET和SPECT成像、医学物理学、结肠癌肿瘤学和临床试验、临床病理学、生物统计学和临床前毒理学等方面的专业知识。

DESCRIPTION (provided by applicant): We hypothesize that a novel capsid incorporation approach will allow comprehensive in vivo monitoring of a cancer retargeted vector, and permit the assessment of our novel vector engineering strategy to enable targeted therapy of disseminated neoplastic disease. By investigating adenovirus (Ad) vector design, we anticipate the new information gained from this initiative will demonstrate the utility of our highly novel imaging approach, and validate the specificity of retargeted tumor transduction. The goal of this application is to develop a multi-functional Ad vector that combines both imaging and targeted therapy. To fulfill this goal, we will combine three unique technologies to advance therapeutic targeting of disseminated neoplastic disease: cancer-specific retargeting, liver-detargeting, and non-invasive imaging. We have developed Ad vectors capable of cell-specific targeting, by incorporating targeting single chain antibodies (scFv) into the viral capsid in combination with hexon modification for liver detargeting and evasion of pre-existing immunity. Our innovative Ad targeting approach provides a novel way to circumvent the problem of structural and biosynthetic incompatibility between Ad and complex targeting ligands such as scFv, and could facilitate Ad targeting to a wide variety of clinically important cell populations using novel targeting ligands including recombinant antibodies and growth factors The goal of this application is to develop a multi-functional Ad vector that combines both imaging and targeted therapy. To fulfill this goal, we will combine three unique technologies to advance therapeutic targeting of disseminated neoplastic disease: cancer-specific retargeting, liver-detargeting, and non-invasive imaging. We have developed Ad vectors capable of cell-specific targeting, by incorporating targeting single chain antibodies (scFv) into the viral capsid in combination with hexon modification for liver detargeting and evasion of pre-existing immunity. Our innovative Ad targeting approach provides a novel way to circumvent the problem of structural and biosynthetic incompatibility between Ad and complex targeting ligands such as scFv, and could facilitate Ad targeting to a wide variety of clinically important cell populations Our innovative use of a structural fusion protein incorporating metallothionein into adenovirus pIX gene provides the non-invasive imaging advantages of detecting physical biodistribution and spread of Ad vectors after administration that is not possible employing a reporter gene. Further, the ability to noninvasively observe Ad function on a whole-body level allows the possibility of detecting virus dissemination outside the tumor site(s) for monitoring clinical safety. By combining these modifications in Ad vector design, we anticipate the new information gained from this initiative will demonstrate the utility of our highly novel imaging approach, and validate the specificity of targeted tumor transduction. This work addresses the significant unmet need of new therapies for advanced stage metastatic cancer. The novel design of the proposed imaging approach is distinct from previously described vector imaging modalities, which have been based exclusively on monitoring the expression of reporter genes. In addition, our combined technologies have broad impact on the diverse field of gene therapy, in which the ability to achieve both cell- specific transduction and monitoring is universally required. To accomplish these goals, we have assembled an outstanding team of investigators with expertise in Ad vector design and construction, adenovirus retargeting approaches, preclinical models using Ad vectors, non-invasive PET and SPECT imaging, medical physics, colon cancer oncology and clinical trials, clinical pathology, biostatistics, and preclinical toxicology.