AntiretroviraI induced Defects in Muscle Protein Synthes

描述（由申请人提供）：本申请是为了响应通知NOT-OD-09-058（NIH宣布恢复法案资金可用于竞争性修订申请）而提交的，并要求根据ARRA 2009进行竞争性补充。我们的母基金提供了关于HIV-1蛋白酶抑制剂如茚地那韦和洛匹那韦（LPV）产生的骨骼肌蛋白质合成减少的机制。该基金的长期目标是阐明LPV和其他蛋白酶抑制剂产生肌肉肌病的机制。在进行母案申请中描述的研究时，我们报告了HIV-1转基因（Tg）大鼠，其是非感染性的并且组成性表达9种病毒包膜蛋白中的7种，表现出肌肉萎缩，而不是归因于热量摄入减少或能量消耗增加。我们现在已经给HIV-1 Tg大鼠注射LPV 3个月，观察到的消瘦程度明显大于未注射LPV的HIV-1 Tg大鼠。此外，我们有证据表明，LPV和HIV-1病毒蛋白表达的共存减少了基础吸收后状态下的肌肉蛋白合成。最后，我们还注意到了一个显着的葡萄糖耐受不良，胰岛素抵抗和异位沉积的甘油三酯在骨骼肌的HIV-1转基因大鼠给予LPV。因此，很明显，在一种或多种病毒蛋白（不是HIV-1病毒本身）和蛋白酶抑制剂之间存在强烈的相互作用，这产生了显著的代谢表型。由于药物-病毒蛋白相互作用，在HIV-1 Tg大鼠中检查蛋白酶抑制剂和其他HAART药物的体内作用似乎提供了一个更具临床相关性的模型。因此，这一竞争性修订的目标是确定LPV给药HIV-1 Tg大鼠产生这种复杂代谢表型的机制-主要关注肌肉中葡萄糖和蛋白质代谢的变化。尽管这些研究是相关的，但它们超出了母公司补助金的直接范围。然而，我们的挑衅性数据值得进一步调查，拟议的研究可以在2年内完成。这一竞争性修订要求为四个具体目的提供资金：1）雇用一名已确定的失业博士后研究员; 2）雇用一名高中化学教师作为暑期实习生; 3）购买极其昂贵的HIV-1转基因大鼠; 4）购买研究员和实习生所需的用品，使其拥有一个富有成效的培训环境，并在两年内成功完成指定的研究。由于拟议的研究不是母申请的一部分，因此没有足够的未支配资金。在母公司申请的最初几年，我们一直非常富有成效，在这个竞争性的修订中提供的资金将大大加快我们的研究，以确定HIV感染者中蛋白酶抑制剂产生的肌肉萎缩和胰岛素抵抗的细胞和分子基础。 公共卫生相关性：在HIV-1感染的患者中，肌肉萎缩和胰岛素抵抗的发生通常与AIDS无关，并且可能是HIV蛋白酶抑制剂药物治疗这些患者的直接结果。我们的研究使用HIV-1转基因大鼠给予常用的蛋白酶抑制剂洛匹那韦，旨在阐明这类药物对肌肉中的翻译效率和胰岛素作用产生负面影响的机制。需要这些数据，以充分发挥这类药物在长期治疗艾滋病毒感染方面的潜力，并避免其可能的缺陷。

DESCRIPTION (provided by applicant): This application is submitted in response to notice NOT-OD-09-058 (NIH Announces the Availability of Recovery Act Funds for Competitive Revision Applications) and requests a competitive supplement under ARRA 2009. Our parent grant has provided mechanism insight regarding the decrease in skeletal muscle protein synthesis produced by HIV-1 protease inhibitors such as indinavir and lopinavir (LPV). The long-term goal of the parent grant is to elucidate mechanisms by which LPV and other protease inhibitors produce muscle myopathy. While conducting the studies described in the parent application, we reported HIV-1 transgenic (Tg) rats, which are noninfectious and constitutively express 7 of 9 viral envelop proteins, exhibit muscle atrophy not attributable to decreased caloric intake or increased energy expenditure. We have now administered LPV to HIV-1 Tg rats for 3 months and the observed wasting was markedly greater than that seen in HIV-1 Tg rats without LPV. Furthermore, we have evidence that the co-existence of LPV and HIV-1 viral protein expression decreases muscle protein synthesis in the basal post-absorptive state. Finally, we also noted a marked glucose intolerance, insulin resistance and ectopic deposition of triglycerides in skeletal muscle of HIV-1 Tg rats administered LPV. Hence, it is clear there is a strong interaction between one or more the viral proteins (not the HIV-1 virus per se) and the protease inhibitor which produces a dramatic metabolic phenotype. It seems likely that examining the in vivo effects of protease inhibitors and other HAART drugs in the HIV-1 Tg rat provides a more clinically relevant model because of the drug-viral protein interaction. Therefore, the goal of this competing revision is to determine the mechanism by which the administration of LPV to HIV-1 Tg rats produces this complex metabolic phenotype - with the primary focus being on the changes in glucose and protein metabolism in muscle. Although related, such studies are beyond the direct scope of the parent grant. However, our provocative data warrant further investigation, and the proposed studies can be completed within a 2-yr period. This competing revision requests funds for four specific purposes: 1) Hiring an identified unemployed postdoctoral fellow; 2) employing a high school chemistry teacher as a summer intern; 3) purchase of HIV-1 Tg rats which are extremely expensive; and 4) purchase of supplies necessary for the fellow and intern to have a productive training environment and to successfully complete the designated studies within 2 years. Because the proposed studies were not part of the parent application sufficient unobligated funds are not available. We have been extremely productive during the initial years of the parent application, and the funds supplied in this competing revision will greatly accelerate our research in identifying the cellular and molecular basis for muscle atrophy and insulin resistance produced by protease inhibitors in HIV-infected individuals. PUBLIC HEALTH RELEVANCE: Muscle wasting and insulin resistance in those patients infected with HIV-1 often occur independent of AIDS and may be a direct result of the treatment of these individuals with HIV protease inhibitor drugs. Our studies using HIV-1 transgenic rats administered the commonly prescribed protease inhibitor lopinavir are designed to elucidate the mechanisms by which this class of drugs negatively impacts translational efficiency and insulin action in muscle. Such data are needed to both realize the full potential and avoid possible pitfalls of this drug class in long-term treatment of HIV infection.