ADENINE DEOXYNUCLEOTIDE METABOLISM AND IMMUNE FUNCTION

腺嘌呤脱氧核苷酸代谢与免疫功能

• 批准号: 2174052
• 负责人: DENNIS CARSON
• 金额: $ 17.95万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 1976
• 资助国家: 美国
• 起止时间: 1976-06-30 至 1996-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2174052
• 关键词: 5' nucleotidase; SCID mouse; adenine nucleotides; apoptosis; cellular pathology; cytotoxicity; deoxyadenosines; disease /disorder model; endonuclease; human subject; immunopharmacology; laboratory rabbit; lymphocyte; lymphocyte proliferation; lymphocytic leukemia; nonhuman therapy evaluation; nucleoside analog; nucleotide metabolism; rheumatoid arthritis; tissue /cell culture

Lymphocytes and monocytes are critical cells in the pathogenesis of immunodeficiency, autoimmune, and certain malignant diseases. It is therefore important to understand the metabolic pathways that regulate lymphocyte and monocyte development and activation under normal and pathologic conditions. This is the long term goal of this research grant application. Previous experiments supported by this research grant have aimed to elucidate the biochemical basis for the association of adenosine deaminase (ADA) deficiency with combined immunodeficiency disease. The results have documented the uniqueness of adenine deoxynucleotide metabolism in lymphocytes and monocytes, and have engendered the development of 2-chlorodeoxyadenosine (CdA). This ADA-resistant analog of deoxyadenosine induces complete remissions in more than 70% of patients with hairy cell leukemia, and has excellent activity in chronic lymphocytic leukemia, and lymphoma. CdA has two biochemical properties that distinguish it from all other nucleoside antimetabolites. First, it is exquisitely toxic to quiescent T and B lymphocytes, that have no replicative DNA synthesis. Second, CdA inhibits the function and survival of monocytes at nanomolar concentrations, that do not affect other phagocytic cells. These biochemical properties suggest that CdA and related compounds should be effective in common autoimmune diseases that are sustained by long-lived memory lymphocytes and activated monocytes. Initial studies in patients with rheumatoid arthritis support this contention. Moreover, CdA and related agents could be useful for the therapy of indolent malignancies that are resistant to cell cycle specific antimetabolites. One main problem that has hindered the widespread testing of CdA in chronic diseases is the need to administer the nucleoside parenterally. In pilot experiments. this problem has been overcome with the synthesis and evaluation of an orally active CdA analog designated CAFdA. Based upon our long-term objective and previous observations, we now aim specifically: (1) To determine how the enzymes that control in vitro sensitivity of quiescent lymphocytes to adenine deoxynucleosides influence the in vivo responsiveness of chronic lymphocytic leukemia cells to CdA therapy. These enzymes include deoxycytidine kinase, cytoplasmic 5'-nucleotidase, and a Ca++/Mg++-dependent endonuclease that mediates apoptosis. (2) To define more clearly the physiologic roles of these three enzymes for the metabolism and function of normal non-dividing lymphocytes and monocytes. (3) To increase the sensitivity of lymphocytes, monocytes, and solid tumors to the inhibitory effects of CdA and CAFdA by interfering with nucleotide dephosphorylation by 5'-nucleotidase, and with cell cycle progression. (4) To prove the in vivo relevance of the results using severe combined immune deficient (SCID) mice reconstituted either with normal human lymphocytes and monocytes, or with cells from patients with rheumatoid arthritis and malignant diseases. (5) Eventually, to initiate new therapies for patients with lymphoproliferative, autoimmune, and malignant diseases.

淋巴细胞和单核细胞是关键细胞 免疫缺陷，自身免疫性和某些恶性疾病。 这是 因此，了解调节的代谢途径很重要 淋巴细胞和单核细胞发育和激活在正常和 病理状况。 这是这项研究赠款的长期目标 应用。这项研究赠款支持的先前实验 旨在阐明腺苷关联的生化基础 脱氨酶（ADA）缺乏伴有免疫缺陷疾病。 这 结果记录了腺嘌呤脱氧核苷酸的独特性 淋巴细胞和单核细胞中的代谢，并引起了 2-氯氧化丙氨酸（CDA）的发展。 这种抗ADA的类似物 脱氧腺苷的诱导诱导超过70％ 毛细胞白血病患者，在慢性病中具有出色的活性 淋巴细胞性白血病和淋巴瘤。 CDA具有两个生化特性 将其与所有其他核苷抗代谢物区分开。 首先，它 对静态T和B淋巴细胞具有精致的毒性 复制DNA合成。 其次，CDA抑制功能，并且 单核细胞在纳摩尔浓度下的存活，不影响 其他吞噬细胞。 这些生化特性表明CDA 相关化合物应在常见的自身免疫性疾病中有效 由长寿命的记忆淋巴细胞维持并激活 单核细胞。 类风湿关节炎患者的初步研究 这个争论。 此外，CDA及相关代理可能对 对细胞周期有抵抗力的顽固性恶性肿瘤的治疗 特定的抗代谢物。 一个阻碍的主要问题 慢性疾病中CDA的广泛测试是需要进行 核苷源自源自源。 在试点实验中。这个问题已经 克服口服活性CDA类似物的合成和评估 指定的CAFDA。 基于我们的长期目标和以前 观察结果，我们现在具体目标：（1）确定酶如何 控制静态淋巴细胞对腺嘌呤的体外敏感性 脱氧核苷影响慢性的体内反应性 淋巴细胞性白血病细胞进行CDA治疗。 这些酶包括 脱氧胞苷激酶，细胞质5'-核苷酸酶和A Ca ++/mg ++ - 介导凋亡的依赖性内切酶。 （2）定义 更清楚地清楚这三种酶的生理作用 正常非分裂淋巴细胞和单核细胞的代谢和功能。 （3）提高淋巴细胞，单核细胞和固体的灵敏度 通过干扰CDA和CAFDA抑制作用的肿瘤 5'-核苷酸酶和细胞周期的核苷酸去磷酸化 进展。 （4）使用 严重的合并免疫缺陷（SCID）小鼠与 正常的人类淋巴细胞和单核细胞，或患有患者的细胞 类风湿关节炎和恶性疾病。 （5）最终，发起 针对淋巴增生性，自身免疫性和 恶性疾病。