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结合方法将允许对癌症重靶向载体进行全面的体内监测，并允许评估我们的新型载体工程策略，以实现播散性肿瘤疾病的靶向治疗。通过对腺病毒（Ad）载体设计的研究，我们预计从这项研究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，并验证重靶向肿瘤转导的特异性。本应用程序的目标是开发一种多功能的广告载体，结合成像和靶向治疗。为了实现这一目标，我们将结合三种独特的技术来推进播散性肿瘤疾病的治疗靶向：癌症特异性重靶向、肝脏去靶向和非侵入性成像。我们已经开发了能够细胞特异性靶向的Ad载体，通过将靶向单链抗体（scFv）结合六邻体修饰结合到病毒衣壳中，以使肝脏脱靶和逃避预先存在的免疫。我们创新的Ad靶向方法提供了一种新的方法，可以避免Ad与复杂靶向配体（如scFv）之间的结构和生物合成不相容问题，并且可以使用新的靶向配体（包括重组抗体和生长因子）促进Ad靶向各种临床重要细胞群。本应用的目标是开发一种多功能Ad载体，结合成像和靶向治疗。为了实现这一目标，我们将结合三种独特的技术来推进播散性肿瘤疾病的治疗靶向：癌症特异性重靶向、肝脏去靶向和非侵入性成像。我们已经开发了能够细胞特异性靶向的Ad载体，通过将靶向单链抗体（scFv）结合六邻体修饰结合到病毒衣壳中，以使肝脏脱靶和逃避预先存在的免疫。我们创新的Ad靶向方法提供了一种新的方法来规避Ad与复杂靶向配体（如scFv）之间的结构和生物合成不相容问题。我们创新地将金属硫蛋白结构融合蛋白结合到腺病毒pIX基因中，提供了检测Ad载体在给药后的物理生物分布和传播的非侵入性成像优势，这是使用报告基因无法实现的。此外，在全身水平上无创观察Ad功能的能力允许检测肿瘤部位外的病毒传播，从而监测临床安全性。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证明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的重大未满足需求。所提出的成像方法的新设计不同于先前描述的矢量成像模式，后者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有着广泛的影响，在这些领域中，实现细胞特异性转导和监测的能力是普遍需要的。为了实现这些目标，我们组建了一支优秀的研究团队，他们在Ad载体的设计和构建、腺病毒重靶向方法、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方面具有专业知识。

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.