SUBSTRATE SUPPLY IN DE NOVO SPHINGOLIPID SYNTHESIS: REGULATION/IMPACT ON CHEMOTH

从头鞘脂合成中的底物供应：对 CHEMOTH 的调节/影响

• 批准号: 8168046
• 负责人: Lauren Ashley Cowart
• 金额: $ 7.3万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8168046
• 关键词: A549; Alternative Splicing; Apoptosis; Apoptotic; Cells; Coenzyme A Ligases; Computer Retrieval of Information on Scientific Projects Database; Data; Fatty Acids; Fatty-acid synthase; Funding; Grant; Heat Stress Disorders; Heating; In Vitro; Institution; Lipids; Mediating; Palmitoyl Coenzyme A; Pathway interactions; Regulation; Research; Research Personnel; Resources; Role; Serine; Source; Sphingolipids; Stimulus; United States National Institutes of Health; Vertebral column; Yeasts; base; cancer cell; chemotherapeutic agent; chemotherapy; enzyme activity; gemcitabine; novel strategies; response; serine palmitoyltransferase; uptake; yeast genetics; A549; Alternative Splicing; Apoptosis; Apoptotic; Cells; Coenzyme A Ligases; Computer Retrieval of Information on Scientific Projects Database; Data; Fatty Acids; Fatty-acid synthase; Funding; Grant; Heat Stress Disorders; Heating; In Vitro; Institution; Lipids; Mediating; Palmitoyl Coenzyme A; Pathway interactions; Regulation; Research; Research Personnel; Resources; Role; Serine; Source; Sphingolipids; Stimulus; United States National Institutes of Health; Vertebral column; Yeasts; base; cancer cell; chemotherapeutic agent; chemotherapy; enzyme activity; gemcitabine; novel strategies; response; serine palmitoyltransferase; uptake; yeast genetics

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Sphingolipids (SLs) mediate cellular activities including apoptosis. The first and rate-limiting step in de novo synthesis of SLs is through serine palmitoyltransferase (SPT), which condenses serine with palmitoyl-CoA, producing a sphingoid base, the backbone of all SLs. Importantly, many stimuli induce de novo synthesis of SLs, including chemotherapy, but the mechanisms are unknown. We have preliminary data that in yeast, heat stress stimulates de novo SL synthesis by stimulating serine uptake from media. Moreover, the fatty acid component for de novo synthesis derives from endogenous synthesis via fatty acid synthase (FAS). Based on these findings, we hypothesize that regulation of substrate supply serves to modulate de novo synthesis of SLs in response to stimuli including chemotherapeutic agents. Therefore, the aims of this proposal are: 1) to determine the mechanisms required for heat stress stimulation of exogenous serine uptake in yeast; 2) to determine the role of FAS and acyl-CoA synthetase in heat-stres induced SPT activity. These 2 aims will be accomplished using a multifaceted approach with yeast genetic sreens, in vitro enzyme activity determinations, and lipid analysis; and 3)to determine the role of substrate supply in cancer cells in response to chemotherapeutic agents. In this aim, we will focus on gemcitabine, which activates the de novo pathway in A549 cells, leading to alternative splicing of pro-apoptotic factors. We hypothesize that substrate availability may regulate gemcitabine-induced de novo SL synthesis, which would allow novel approaches to chemotherapy.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 鞘脂（SLS）介导包括细胞凋亡的细胞活性。 SLS从头合成的第一个且限制的步骤是通过丝氨酸棕榈酰转移酶（SPT），它与棕榈酰-COA凝结，产生鞘脂基碱，这是所有SLS的骨干。 重要的是，许多刺激诱导了SLS的从头合成，包括化学疗法，但这些机制尚不清楚。 我们拥有初步数据，这些数据在酵母中，热应力通过刺激培养基的丝氨酸摄取来刺激从头综合。 此外，从头合成的脂肪酸成分通过脂肪酸合酶（FAS）源自内源性合成。 基于这些发现，我们假设底物供应的调节可用于调节SLS的从头合成，以响应包括化学治疗剂在内的刺激。因此，该提案的目的是：1）确定酵母中外源丝氨酸摄取所需的机制； 2）确定FAS和酰基-COA合成酶在热元诱导的SPT活性中的作用。 这2个目标将使用酵母遗传性的多方面方法，体外酶活性测定和脂质分析来实现。 3）确定底物供应在癌细胞中对化学治疗剂的作用。在此目标中，我们将专注于吉西他滨，该吉西他滨在A549细胞中激活从头途径，从而导致促凋亡因子的替代剪接。我们假设底物的可用性可能调节吉西他滨诱导的从头合成，这将允许新颖的化学疗法方法。