Immunotherapy of CLL with lentiviral expressed CD40-ligand

项目摘要/摘要：尽管近年来取得了一些进展，但作为最常见的白血病，CLL在很大程度上仍然是一种无法治愈的疾病。该项目的目标是阐明慢病毒载体（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细胞刺激物。这种使用LV-CD154的方法有望优于使用腺病毒载体的现有免疫疗法，并将成为未来CLL和潜在的其他血液系统恶性肿瘤改进免疫疗法的基础。在目标1中，我们将开发表达cd40配体的SIN慢病毒载体，以及表达GFP的对照载体。所有载体均含有果蝇dNK-B10基因，采用E2A裂解肽序列，提高载体的安全性。LV将按照标准程序制造，并在放行前进行质量控制测试。在Aim 2中，我们将测试SIN.LV的转导效率。CD154和sin。lv。此外，我们将评估转染基因的表达持续时间，并测试可溶性CD154靶向细胞刺激与细胞结合CD154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细胞中的功效。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III期，该疗法将被商业化用于CLL和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。所产生的结果将为开发更好的CLL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：尽管近年来取得了一些进展，但作为最常见的白血病，慢性淋巴细胞白血病（CLL）在很大程度上仍然是一种无法治愈的疾病。现代治疗方案包括嘌呤类似物、单克隆抗体、移植策略和免疫疗法等新药。在这个提议中，我们期望产生新的CLL免疫疗法，可能会迅速扩展到其他血液系统恶性肿瘤的治疗。这种疗法对白血病和淋巴瘤患者具有重要意义，最终对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.