The Dichotomy of Alk1 and Alk5 Signaling Pathways in Vascular Response to Injury

Alk1 和 Alk5 信号通路在血管损伤反应中的二分法

基本信息

批准号：
8847768
负责人：
Zhihua Jiang
金额：
$ 31.36万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-07-15 至 2017-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8847768
关键词：
Adult Anti-Inflammatory Agents Anti-inflammatory Attenuated Autologous Automobile Driving Behavior Biological Models Biology Blood Vessels Bypass CCL2 gene Cell Culture Techniques Cells Cellular biology Cessation of life Clinical Clinical Trials Complex Cre-LoxP Data Development Endothelial Cells Endothelium Equilibrium Evaluation Failure Fibrosis Genes Genetic Genetic Recombination Healed Human Hyperplasia Implant In Vitro Inflammatory Interleukin-6 Knowledge Laboratories Limb structure Medial Mediating Mediator of activation protein Modeling Morphology Mus Myosin Heavy Chains NF-kappa B Nature Organ Outcome Pathology Patients Performance Pharmacologic Substance Phenotype Population Process Production Property Recovery of Function Reporting Role Secondary to Signal Pathway Signal Transduction Small Interfering RNA Smooth Muscle Myocytes Specificity Stenosis System Tamoxifen Testing Therapeutic Transforming Growth Factors Translations United States Veins Work angiogenesis cancer therapy cell type clinical application clinically relevant clinically significant cytokine genetic approach graft failure graft healing healing improved in vivo inhibitor/antagonist insight leukemic stem cell migration monolayer novel novel strategies programs promoter reagent testing receptor recombinase repaired response response to injury success therapeutic effectiveness tool treatment strategy

项目摘要

DESCRIPTION (provided by applicant): Over a half million of autologous vein grafts are implanted annually in the United States. However, 30-60% of the grafts fails or develops a clinically significant stenosis within the first year, causing limb loss and death. The primary cause for early vein graft failure has been identified as neointimal hyperplasia (NIH) and compelling evidence has demonstrated that TGF-ß is a driving factor for this early failure. Unfortunately, non-selective blockade of the broad TGF-ßactivities has yielded limited success in attenuating neointimal hyperplasia formation, suggesting inhibition of the specific TGF-ß activities is required. A primary mechanism that dictates TGF-ß specificity is the activation of its type I receptors Alk1 or Alk5. Although Alk1 is expressed at very low level in mature endothelium (ECs) and medial smooth muscle cells (SMCs), existing evidence suggests that Alk1 is induced in ECs and neointimal SMCs during vein graft adaptation. Recent studies for angiogenesis and other pathologies have led to an emergence of new understanding, wherein TGF-ß signals through Alk1 and Alk5 to initiate opposing effects on regulating cellular biology. We therefore hypothesize that the response of the vascular wall to TGF-ß relies on the balance between Alk1- and Alk5- signaling in both ECs and SMCs. Insult to the vein graft wall tips the balance in both cell types towards Alk5 signaling that in turn inhibits the functional recovery of ECs and upholds an inflammatory/synthetic phenotype for SMCs, driving progressive NIH. Selectively blocking Alk5 signaling to restore this balance will improve the healing response and inhibit NIH. To test this hypothesis, this project aims to: 1) Define the role of Alk1 and Alk5 signaling in SMCs in regulating the phenotype of neointimal SMCs and vein graft morphology via a validated murine vein graft model and primary neointimal SMC culture; 2) Evaluate the impact of the competing Alk1 and Alk5 signaling in ECs on functional recovery of the repopulated EC monolayer, modulation of neoSMC phenotype, and the resultant vein graft morphology; and 3) Examine the therapeutic effectiveness of siRNA and pharmaceutical inhibition of Alk1 or Alk5 signaling on the development of NIH in murine and human vein grafts. The CreloxP system will be utilized to induce selective deletion of Alk1 or Alk5 in ECs or SMCs in adult mice, so that vein grafts with and without EC or SMC specific Alk1 or Alk5 can be created for the evaluation of the vein graft morphology, the repair of the EC monolayer, and the inflammatory phenotype of neointimal SMCs. To facilitate the clinical translation of the new knowledge generated with these genetic approaches, specific siRNA and novel pharmacological inhibitors will be applied to inhibit Alk1 and Alk5 signaling pathways in both murine and ex vivo human vein grafts. The therapeutic effectiveness of these approaches will then be evaluated using both morphologic (e.g. NIH volume) and biologic (e.g. phenotypic properties of the neointimal cells) endpoints. Completion of these aims will not only provide new insights into the fundamentals of TGF-ß biology, but also generate novel strategies to manipulate complex biologic processes such as vein graft wall adaptation.

描述（由申请人提供）：美国每年植入超过 50 万个自体静脉移植物，但是，30-60% 的移植物在第一年内失败或出现临床上显着的狭窄，导致肢体丧失和死亡。早期静脉移植失败的主要原因已被确定为新生内膜增生 (NIH)，令人信服的证据表明 TGF-β 是这种早期失败的驱动因素，不幸的是，非选择性阻断。广泛的 TGF-β 活性在减弱新内膜增生形成方面取得了有限的成功，表明需要抑制特定的 TGF-β 活性，尽管 Alk1 被表达，但决定 TGF-β 特异性的主要机制是激活其 I 型受体 Alk1 或 Alk5。在成熟内皮细胞 (EC) 和内侧平滑肌细胞 (SMC) 中水平非常低，现有证据表明 Alk1 在 EC 中被诱导，并且最近对血管生成和其他病理学的研究产生了新的认识，因此 TGF-β 通过 Alk1 和 Alk5 发出信号来启动对调节细胞生物学的相反作用。血管壁至 TGF-β 依赖于 EC 和 SMC 中 Alk1- 和 Alk5- 信号传导之间的平衡。转抑制 EC 的功能恢复并维持 SMC 的炎症/合成表型，从而驱动进行性 NIH。选择性阻断 Alk5 信号传导以恢复这种平衡将改善愈合反应并抑制 NIH。为了检验这一假设，该项目旨在：1)通过经过验证的鼠静脉移植模型和原代新内膜 SMC 培养，定义 SMC 中 Alk1 和 Alk5 信号传导在调节新内膜 SMC 表型和静脉移植形态方面的作用； 2) 评估 EC 中竞争性 Alk1 和 Alk5 信号传导对重新填充的 EC 单层的功能恢复、neoSMC 表型的调节以及由此产生的静脉移植物形态的影响；3) 检查 siRNA 的治疗效果和 Alk1 或的药物抑制作用Alk5 信号对小鼠和人静脉移植物中 NIH 发展的影响 CreloxP 系统将用于诱导 EC 或中 Alk1 或 Alk5 的选择性删除。成年小鼠体内的 SMC，可以制作带有或不带有 EC 或 SMC 特异性 Alk1 或 Alk5 的静脉移植物，用于评估静脉移植物的形态、EC 单层的修复以及新生内膜 SMC 的炎症表型，以方便临床。通过这些遗传方法产生的新知识的转化，特异性 siRNA 和新型药理学抑制剂将应用于抑制小鼠和离体人静脉移植物中的 Alk1 和 Alk5 信号通路。然后将使用形态学（例如 NIH 体积）和生物学（例如新内膜细胞的表型特性）终点来评估方法。完成这些目标不仅将为 TGF-ß 生物学的基础提供新的见解，而且还会产生新的策略。操纵复杂的生物过程，例如静脉移植壁适应。

项目成果

期刊论文数量（7）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Smooth muscle cell-specific Tgfbr1 deficiency promotes aortic aneurysm formation by stimulating multiple signaling events.

平滑肌细胞特异性 Tgfbr1 缺陷通过刺激多种信号传导事件促进主动脉瘤形成。

DOI：
发表时间：
2016
期刊：
影响因子：
4.6
作者：
Yang, Pu;Schmit, Bradley M;Fu, Chunhua;DeSart, Kenneth;Oh, S Paul;Berceli, Scott A;Jiang, Zhihua
通讯作者：
Jiang, Zhihua

An X-linked Myh11-CreERT2 mouse line resulting from Y to X chromosome-translocation of the Cre allele.

X 连锁 Myh11-CreERT2 小鼠系由 Cre 等位基因的 Y 染色体易位至 X 染色体而产生。

DOI：
发表时间：
2017-09
期刊：
影响因子：
0
作者：
Liao, Mingmei;Zhou, Junmei;Wang, Fen;Ali, Yasmin H;Chan, Kelvin L;Zou, Fei;Offermanns, Stefan;Jiang, Zhisheng;Jiang, Zhihua
通讯作者：
Jiang, Zhihua

Cyclophilin A contributes to aortopathy induced by postnatal loss of smooth muscle TGFBR1.

亲环蛋白 A 会导致出生后平滑肌 TGFBR1 缺失引起的主动脉病。

DOI：
发表时间：
2019
期刊：
影响因子：
0
作者：
Zhou, Guannan;Liao, Mingmei;Wang, Fen;Qi, Xiaoyan;Yang, Pu;Berceli, Scott A;Sharma, Ashish K;Upchurch Jr, Gilbert R;Jiang, Zhihua
通讯作者：
Jiang, Zhihua

Hypertension overrides the protective effect of female hormones on the development of aortic aneurysm secondary to Alk5 deficiency via ERK activation.

高血压超越了女性激素对 Alk5 缺乏通过 ERK 激活继发的主动脉瘤发展的保护作用。