Molecular Regulation of Folate and Antifolate Transport

叶酸和抗叶酸转运的分子调节

基本信息

批准号：
10652998
负责人：
Zhanjun Hou
金额：
$ 29.26万
依托单位：
WAYNE STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1993
资助国家：
美国
起止时间：
1993-02-12 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10652998
关键词：
Acids Alimta Anabolism Anatomy Anions Antineoplastic Agents Biological Biology Cancer Etiology Carbon Cell Line Cell Survival Cell membrane Cells Circulation Clinical Complex Coupled Cytosol Diffusion Disease Drug resistance Epithelial Cells Epithelial ovarian cancer Epithelium FRAP1 gene Folic Acid Folic Acid Antagonists Folic Acid Deficiency Formates Generations Genetic Transcription Glutathione Glycine Glycine Hydroxymethyltransferase Goals Grant Human Human Biology Hydroxymethyltransferases Hypoxia Immunohistochemistry Leukemic Cell Lung Adenocarcinoma Malabsorption Syndromes Malignant Neoplasms Malignant Pleural Mesothelioma Mammalian Cell Mediating Membrane Membrane Transport Proteins Metabolic Pathway Metabolism Methionine Methotrexate Mitochondria Molecular Mutation Normal Cell Orphan Drugs Ovarian Pancreas Pancreatic Adenocarcinoma Pathway interactions Patients Pemetrexed Pharmacodynamics Physiological Physiology Protons Publishing Purine Nucleotides Purines Raltitrexed Reactive Oxygen Species Regulation Reporting Research Resistance Ribonucleotides Role SLC19A1 gene Series Serine Signal Transduction Specimen Structure-Activity Relationship System Tetrahydrofolates Therapeutic Time Tissues Tumor Cell Line Tumor stage Upper digestive tract structure absorption antitumor agent antitumor drug autosome clinical translation cytotoxic design differential expression folate-binding protein glycine amide hereditary folate malabsorption human disease hydrophilicity in vivo inhibitor neoplastic cell novel novel strategies novel therapeutics protein protein interaction targeted agent targeted treatment thymidylate tumor tumor microenvironment tumor progression tumor xenograft uptake

项目摘要

ABSTRACT Folates are anionic molecules that cross biological membranes poorly by diffusion. Folate uptake is principally mediated by the reduced folate carrier (RFC; SLC19A1) and the proton-coupled folate transporter (PCFT; SLC46A1). Upon internalization, folates facilitate one-carbon (C1) metabolism, leading to synthesis of glycine, serine and methionine, and purine nucleotides and thymidylate. C1 metabolism encompasses cytosolic and mitochondrial pathways connected by an interchange between serine, glycine and formate. The ubiquitously expressed RFC is the major membrane transporter for folates in cells and tissues. RFC is also an important transporter of clinically used C1 inhibitors (e.g., pemetrexed) for cancer, as well as other indications, and loss of RFC is associated with drug resistance. PCFT mediates folate absorption in the upper gastrointestinal tract. PCFT levels in other tissues are generally modest. Unlike RFC, PCFT transport is optimal at acidic pH, approximating the tumor microenvironment. PCFT is widely expressed in human tumor cell lines and primary specimens. We discovered novel cytotoxic PCFT-targeted C1 inhibitors for cancer and established a comprehensive structure-activity relationship for PCFT that is distinct from RFC. Novel pyrrolopyrimidine compounds (AGF94 & AGF347) showed potent anti-proliferative activities toward PCFT-expressing tumors that were augmented at acid pH. Following internalization, AGF94 inhibited de novo purine (DNP) biosyn- thesis at β-glycinamide ribonucleotide formyltransferase (GARFTase), whereas AGF347 inhibited mito- chondrial C1 metabolism at serine hydroxymethyltransferase 2 (SHMT2), with additional effects on C1 metabolism in the cytosol (DNP biosynthesis at GARFTase and 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase, and at SHMT1). Both inhibitors depleted ATP; AGF347 depleted glycine and cytosolic C1 pools, with downstream effects on glutathione and levels of reactive oxygen species, and on mTOR signaling. AGF94 and AGF347 showed promising in vivo efficacies toward early and upstage tumor xeno- grafts. We posit that our PCFT-targeted agents offer an entirely new approach for treating cancer. In this R01 renewal, we explore the unique biology of the facilitative folate transporters and C1 metabolism, with a goal of further optimizing therapeutic applications of our novel agents. We propose in Aim 1 to characterize the cellular pharmacodynamics and molecular regulation of PCFT in relation to PCFT-targeted therapies, including transcriptional mechanisms and the role of protein-protein interactions in regulating PCFT. In Aim 2, we will characterize the cellular pharmacodynamics of mitochondrial C1 inhibitors including their transport and metabolism. An important focus of both Aims 1 and 2 will be on the role of the tumor microenvironment, including the impact of hypoxia and acid pH on anti-tumor drug biology and efficacy of these series. Our proposed studies are distinctive for their novelty and focus on clinical translation.

抽象的叶状是阴离子分子，通过扩散较差的生物膜横穿生物膜。叶酸的摄取主要是由还原的叶酸载体（RFC； SLC19A1）和质子偶联的叶酸转运蛋白（PCFT; SLC46A1）。内在化后，叶酸盐促进了一种碳（C1）代谢，从而导致甘氨酸的合成，丝氨酸和蛋氨酸，嘌呤核苷酸和胸苷酸。 C1代谢包括胞质和通过丝氨酸，甘氨酸和形式之间的互换连接的线粒体途径。无处不在表达的RFC是细胞和组织中叶酸的主要膜转运蛋白。 RFC也很重要用于癌症的临床使用的C1抑制剂（例如，Pemetrexed）的转运蛋白以及其他适应症以及丧失 RFC与耐药性有关。 PCFT介导上胃肠道的叶酸滥用。其他组织中的PCFT水平通常适中。与RFC不同，PCFT转运在酸性pH下是最佳的近似肿瘤微环境。 PCFT在人类肿瘤细胞系和原发性中广泛表达标本。我们发现了新型的细胞毒性PCFT靶向癌症的C1抑制剂，并建立了 PCFT的综合结构活性关系与RFC不同。新型吡咯吡汀化合物（AGF94和AGF347）显示了针对表达PCFT的肿瘤的有效抗增殖活动在酸性pH下增加了。内在化后，AGF94抑制了从头嘌呤（DNP）Biosyn- 在β-糖苷酰胺核糖核苷酸甲基转移酶（GARFTase）的论文，而AGF347抑制了mito- 丝氨酸羟基转移酶2（SHMT2）的软骨C1代谢，对C1有其他影响细胞质中的代谢（Garftase和5-氨基咪唑-4-羧酰胺的DNP生物合成核糖核苷酸甲基转移酶，在SHMT1）。两种抑制剂都耗尽了ATP； AGF347耗尽甘氨酸和胞质C1池，对谷胱甘肽和活性氧的水平以及MTOR具有下游影响信号。 AGF94和AGF347表现出对早期和升级肿瘤Xeno-的体内效率的希望移植物。我们认为，我们的PCFT靶向剂为治疗癌症提供了一种全新的方法。在此R01中更新，我们探索了最喜欢的叶酸转运蛋白和C1代谢的独特生物学进一步优化我们新型药物的治疗应用。我们在AIM 1中提出以表征 PCFT的细胞药效学和分子调节与PCFT靶向疗法有关，包括转录机制和蛋白质 - 蛋白质相互作用在调节PCFT中的作用。在AIM 2中，我们将表征线粒体C1抑制剂（包括其运输和）的细胞药效学代谢。目标1和2的一个重要重点将放在肿瘤微环境的作用上，包括缺氧和酸pH对这些系列的抗肿瘤药物生物学的影响。我们的拟议的研究对于它们的新颖性和专注于临床翻译而言是独特的。

项目成果

期刊论文数量（10）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Biology of the major facilitative folate transporters SLC19A1 and SLC46A1.

DOI：
10.1016/b978-0-12-800223-0.00004-9
发表时间：
2014
期刊：
CURRENT TOPICS IN MEMBRANES
影响因子：
0
作者：
Hou, Zhanjun;Matherly, Larry H.
通讯作者：
Matherly, Larry H.

The evolving biology of the proton-coupled folate transporter: New insights into regulation, structure, and mechanism.

DOI：
10.1096/fj.202101704r
发表时间：
2022-03
期刊：
FASEB journal : official publication of the Federation of American Societies for Experimental Biology
影响因子：
0
作者：
Hou Z;Gangjee A;Matherly LH
通讯作者：
Matherly LH

Substrate-specific binding and conformational changes involving Ser313 and transmembrane domain 8 of the human reduced folate carrier, as determined by site-directed mutagenesis and protein cross-linking.

通过定点诱变和蛋白质交联测定涉及人还原叶酸载体的 Ser313 和跨膜结构域 8 的底物特异性结合和构象变化。

DOI：
10.1042/bj20100181
发表时间：
2010
期刊：
The Biochemical journal
影响因子：
0
作者：
Hou,Zhanjun;Wu,Jianmei;Ye,Jun;Cherian,Christina;Matherly,LarryH
通讯作者：
Matherly,LarryH

Discovery of Tumor-Targeted 6-Methyl Substituted Pemetrexed and Related Antifolates with Selective Loss of RFC Transport.

DOI：
10.1021/acsmedchemlett.3c00326
发表时间：
2023-12-14
期刊：
ACS MEDICINAL CHEMISTRY LETTERS
影响因子：
4.2
作者：
Kaku, Krishna;Ravindra, Manasa P.;Tong, Nian;Choudhary, Shruti;Li, Xinxin;Yu, Jianming;Karim, Mohammad;Brzezinski, Madelyn;O'Connor, Carrie;Hou, Zhanjun;Matherly, Larry H.;Gangjee, Aleem
通讯作者：
Gangjee, Aleem

Effects of 5' untranslated region diversity on the posttranscriptional regulation of the human reduced folate carrier.

5非翻译区多样性对人类还原叶酸载体转录后调节的影响。