Immunotherapy of CLL with lentiviral expressed CD40-ligand

描述(申请人提供)：项目摘要/摘要：尽管最近取得了进展，但作为最常见的白血病，CLL仍然是一种基本上无法治愈的疾病。本项目的目的是阐明慢病毒载体(LV)为CD40配体(CD154)提供高效和临床可行的递送系统的能力。CD154是慢性淋巴细胞白血病(CLL)中非常著名的T细胞刺激因子。这种使用LV-CD154的方法有望优于现有的使用腺病毒载体的免疫疗法，并将为未来改进CLL和潜在的其他血液系统恶性肿瘤的免疫疗法奠定基础。在目标1中，我们将构建表达CD40配体的SIN慢病毒载体和表达GFP的对照载体。所有载体都将包含果蝇DNK-B10基因，使用E2a裂解肽序列，这将增加载体的安全性。LV将根据标准程序和质量控制进行制造，并在发布进行测试之前进行测试。在目标2中，我们将测试SIN.LV.CD154和SIN.LV.GFP在原代CLL细胞和不同低度恶性淋巴瘤细胞系(CLL/SLL，MCL和FL)的体外转导效率。此外，我们还将评估转基因表达的持续时间，并测试可溶性CD154与细胞结合CD154靶向刺激细胞的相对效力。在第一阶段，我们建议产生含有CD154有效载荷的SIN-LV，并研究其在CLL细胞中的有效性。最终，在第二阶段，SIN-LV-.CD154将在临床前和临床环境中进行测试。在第三阶段，这种疗法将用于慢性淋巴细胞性白血病以及可能的其他血液系统恶性肿瘤，如套细胞淋巴瘤和急性髓系白血病。这些结果将为开发更好的治疗CLL的新疗法提供基础，并将极大地挑战现有的使用腺病毒载体的免疫疗法，这些疗法大多不足以提供稳定和成功的长期治疗活性。项目简介：尽管最近取得了进展，但慢性淋巴细胞白血病(CLL)作为最常见的白血病仍然是一种基本上无法治愈的疾病。现代治疗选择包括新型药物，如嘌呤类似物、单抗、移植策略和免疫疗法。在这项提议中，我们希望产生新的CLL免疫疗法，这种疗法很可能迅速扩展到其他血液系统恶性肿瘤的治疗。这种疗法将对白血病和淋巴瘤患者产生重大影响，最终对一般公众健康产生积极影响。

DESCRIPTION (provided by applicant): Project Summary/Abstract: Despite recent advances, CLL as the most common leukemia remains a largely incurable disease. The goal of the proposed project is to elucidate the ability of the lentiviral vector (LV) to provide efficient and clinically feasible delivery system for CD40-ligand (CD154), a very well known stimulator of T cells in chronic lymphocytic leukemia (CLL). This approach using LV-CD154 is expected to be superior to existing immune therapies using adenoviral vectors and will be the basis for a future improved immune therapy for CLL and potentially other hematological malignancies. In Aim 1, we will develop SIN lentiviral vector that expresses CD40-ligand, as well as control vector expressing GFP. All vectors will contain the Drosophila melanogaster dNK-B10 gene using the E2A cleavage peptide sequence which will increase vector's safety. LV will be manufactured according to standard procedures and quality control tested prior to release for testing. In Aim 2, we will test the transduction efficiency of SIN.LV.CD154 and SIN.LV. GFP in vitro using primary CLL cells, and cell lines from different low grade lymphomas (CLL/SLL, MCL and FL) In addition, we will assess duration of expression of the transfected gene and test the relative potency of targeted cell stimulation with soluble CD154 versus cell bound CD154. During Phase I, we propose to generate SIN-LV containing CD154 payload and to investigate its efficacy in CLL cells. Ultimately, during Phase II, the SIN-LV-.CD154 will be tested in preclinical and clinical settings. During Phase III, this therapy will be commercialized for CLL and possibly other hematological malignancies like mantle cell lymphoma and acute myeloid leukemia. The generated results will provide a basis for the development of a better and novel therapy for CLL and will significantly challenge the existing immune therapies using adenoviral vectors, mostly inadequate for providing stable and successful long term therapeutic activity. Project Narrative: Despite recent advances, chronic lymphocytic leukemia(CLL) as the most common leukemia remains a largely incurable disease. Modern treatment options include novel drugs like purine analogues, monoclonal antibodies, transplantation strategy and immunotherapy. In this proposal, we expect to generate novel immune therapy of CLL that is likely to quickly expand to treatment of other hematological malignancies. This therapy will have significant relevance for leukemia and lymphoma patients, ultimately positively impacting public health in general.