Restoration of Fecal Continence in Aging IAS

老年 IAS 患者大便失禁的恢复

基本信息

批准号：
8214650
负责人：
KHALIL N BITAR
金额：
$ 30.57万
依托单位：
WAKE FOREST UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-02-01 至 2014-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8214650
关键词：
3-Dimensional Acetylcholine Actins Adrenergic Agents Adult Affect Age Aging Aliquot Antibodies Anus Applications Grants Area Biomedical Engineering Budgets Canis familiaris Caveolae Caveolins Cell Aging Cells Circular layer of muscularis propria of anal canal Colon Complementary DNA Complex Constipation Contractile Proteins Contracts Cyclic AMP Cytoskeletal Modeling Cytoskeleton Data Development Dominant-Negative Mutation Dose Down-Regulation Elderly Enteric Nervous System Exhibits Extravasation Fecal Incontinence Feces Figs - dietary Functional disorder Gastrointestinal Contents Gastrointestinal Sphincter Generations HSPB1 gene Hand Hormonal Human In Vitro Interphase Cell Investigation Lead Length Lipids Maintenance Mechanics Mediator of activation protein Membrane Membrane Microdomains Methods Modeling Molecular Motor Movement Muscle Muscle Cells Muscle Contraction Muscle function Nature Nerve Neurotransmitters Outcome Phosphorylation Physiological Play Principal Investigator Property Protein Isoforms Quality of life Rattus Rectum Relaxation Rest Role Signal Transduction Skeletal Muscle Smooth Muscle Smooth Muscle Myocytes Sphincter Sucrose Time Tissue Engineering Work adrenergic age effect age related aged caveolin 1 cell age cholinergic cost density in vitro Model in vivo loss of function novel overexpression pressure programs research study response restoration

项目摘要

Principal Investigator/Program Director (Last, first, middle): Bitar, Khalil, N RESEARCH & RELATED Other Project Information 1. * Are Human Subjects Involved? m Yes l No 1.a. If YES to Human Subjects Is the IRB review Pending? m Yes m No IRB Approval Date: Exemption Number: 1 2 3 4 5 6 Human Subject Assurance Number 2. * Are Vertebrate Animals Used? l Yes m No 2.a. If YES to Vertebrate Animals Is the IACUC review Pending? m Yes l No IACUC Approval Date: 11-29-2006 Animal Welfare Assurance Number A3114-01 3. * Is proprietary/privileged information m Yes l No included in the application? 4.a.* Does this project have an actual or potential impact on m Yes l No the environment? 4.b. If yes, please explain: 4.c. If this project has an actual or potential impact on the environment, has an exemption been authorized or an environmental assessment (EA) or environmental impact statement (EIS) been performed? m Yes m No 4.d. If yes, please explain: 5.a.* Does this project involve activities outside the U.S. or m Yes l No partnership with International Collaborators? 5.b. If yes, identify countries: 5.c. Optional Explanation: 6. * Project Summary/Abstract 7519-PROJECTABSTRACT.pdf Mime Type: application/pdf 7. * Project Narrative 8426-PROJECT_NARRATIVE.pdf Mime Type: application/pdf 8. Bibliography & References Cited 5924-REFERENCESCITED.pdf Mime Type: application/pdf 9. Facilities & Other Resources 3457-RESOURCES.pdf Mime Type: application/pdf 10. Equipment 7977-MAJOR_EQUIPMENT.pdf Mime Type: application/pdf Tracking Number: Other Information Page 5 OMB Number: 4040-0001 Expiration Date: 04/30/2008 Principal Investigator/Program Director (Last, first, middle): Bitar, Khalil, N ABSTRACT Little is known about the mechanisms or pathophysiology responsible for age-related decline of internal anal sphincter (IAS) function. Decreased mechanical efficiency of smooth muscle of the IAS results in decreased closure pressure of the sphincter, thus greatly contributing to fecal incontinence which is disproportionately prevalent in the elderly. Recent advances in tissue engineering provide us with an excellent in vitro model mimicking in vivo function to study the effects of aging on the molecular mechanisms of the IAS smooth muscle. We have for the first time, bioengineered three- dimensional (3-D) rings from isolated smooth muscle cells from human IAS. These rings developed tone, responded to acetylcholine in a dose-dependent manner and relaxed upon the exogenous addition of the relaxant mediator 8-Br-cAMP. Preliminary results from Human IAS and Human circular colonic smooth muscle cells (CSMC) indicate: 1) Sequestration of RhoA, phospho-PKC¿ (S657) and HSP27 only in the caveolin-rich lipid raft microdomains of IAS cells at rest and not CSMC; 2) a greater expression of HSP27, RhoA, PKC¿ and phospho CPI-17 in Human IAS cells vs. CSMC. Rings from old Rat IAS cells showed decreased contractile response (maximal force generation ¿N and time-to-peak response) when compared to IAS rings from adult rats that correlated with decreased HSP27 phosphorylation. Reduced phosphorylation of HSP27 affects actin cytoskeleton stability leading to disturbed caveolae formation. Overexpression of phosphomimic-HSP27 in old IAS smooth muscle cells exhibited increased association of PKC¿ and HSP27 with caveolin-1, and also reinstated the magnitude of force generated and the time-to-peak contraction in IAS rings bioengineered from these cells. The specific aims of this grant proposal are: 1) Develop a 3-D physiological model of the IAS bioengineered in vitro from cells isolated from the IAS of human and of adult and aged rats, and examine the effect of aging on the molecular mechanisms of tonic IAS smooth muscle contraction 2) Study the role of phosphorylated-HSP27 in age-related decline of IAS smooth muscle function, and 3) Examine the reinstatement of physiological contractile function in 3-D IAS rings bioengineered from IAS smooth muscle cells transfected with phosphomimic-HSP27 cDNA. Project Description Page 6

首席研究员/计划主任（最后，第一，中间）：比特尔，哈利勒，n 研究及相关其他项目信息 1。 *人类受试者是否涉及？ M是L否 1.A.如果是人类主题 IRB审查正在待处理吗？ M是M否 IRB批准日期：豁免号：1 2 3 4 5 6 人类主题保证号 2。 *使用脊椎动物吗？ L是M否 2.A.如果是脊椎动物 IACUC审查尚待审查吗？ M是L否 IACUC批准日期：11-29-2006 动物福利保证号A3114-01 3。 *是专有/特权信息m是l否包括在应用程序中？ 4.A.*该项目对M是L否具有实际或潜在的影响环境？ 4.B.如果是，请说明： 4.C.如果该项目对环境有实际或潜在的影响，则授权豁免或环境评估（EA）或进行了环境影响声明（EIS）？ M是M否 4.D.如果是，请说明： 5.A.*该项目是否涉及美国以外的活动或M是L否与国际合作者合作？ 5.B.如果是，请确定国家： 5.C.可选解释： 6。 *项目摘要/摘要7519-projectabstract.pdf Mime类型：应用程序/PDF 7。 *项目叙述8426-project_narrative.pdf Mime类型：应用程序/PDF 8。引用的参考书目和参考文献5924-参考。PDFMIME类型：应用程序/PDF 9。设施和其他资源3457-Resources.pdf Mime类型：应用程序/PDF 10。设备7977-Major_equipment.pdf Mime类型：应用程序/PDF 跟踪号：其他信息第5页OMB编号：4040-0001 到期日期：2008年4月30日首席研究员/计划主任（最后，第一，中间）：比特尔，哈利勒，n 抽象的关于负责年龄有关的机制或病理生理学知之甚少内部肛门括约肌（IAS）功能的下降。降低机械效率 IAS的平滑肌导致括约肌的闭合压力提高，因此极大地导致粪便尿失禁，这在老年。组织工程的最新进展为我们提供了极好的体外模型模拟体内功能以研究衰老对分子的影响 IAS平滑肌的机制。我们第一次有生物工程三来自人类IAS的分离平滑肌细胞的尺寸（3-D）环。这些戒指开发的音调，以剂量依赖性的方式对乙酰胆碱反应并放松外源添加放松介质8-Br训练训练。初步结果来自人类IAS和人圆的结肠平滑肌细胞（CSMC）表示：1）仅在富含小窝蛋白的脂质中，RhoA，Phosho-Pkc¿（S657）和Hsp27的固相 IAS细胞在静止而不是CSMC时的筏微区； 2）HSP27的更大表达，人IAS细胞中的RhoA，PKCC和Phospho CPI-17与CSMC。老鼠的戒指 IAS细胞显示收缩反应降低（最大力产生�N和与与成年大鼠相关的IAS环相比 HSP27磷酸化降低。 HSP27的磷酸化降低会影响肌动蛋白细胞骨架稳定性导致分布式小窝形成。过表达旧IAS平滑肌细胞中的磷光-HSP27表现出增加 pkc¿和hsp27带有小窝蛋白-1，还恢复了产生的力的大小以及来自这些细胞生物工程的IAS环的峰值收缩。这该赠款提案的具体目的是：1）开发IAS的3D物理模型从从人类和成人和老年人的IAS分离的细胞中进行生物工程大鼠，并检查衰老对滋补IAS平滑分子机制的影响肌肉收缩2）研究磷酸化-HSP27在年龄相关下降的作用 IAS平滑肌功能，3）检查生理的恢复原状从IAS平滑肌细胞生物工程的3-D IAS环的收缩功能用磷酸HSP27 cDNA转染。项目说明第6页

项目成果

期刊论文数量（12）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（3）

Successful implantation of an engineered tubular neuromuscular tissue composed of human cells and chitosan scaffold.

成功植入由人体细胞和壳聚糖支架组成的工程管状神经肌肉组织。

DOI：
10.1016/j.surg.2015.05.009
发表时间：
2015
期刊：
Surgery
影响因子：
3.8
作者：
Zakhem,Elie;Elbahrawy,Mostafa;Orlando,Giuseppe;Bitar,KhalilN
通讯作者：
Bitar,KhalilN

Enteric neural differentiation in innervated, physiologically functional, smooth muscle constructs is modulated by bone morphogenic protein 2 secreted by sphincteric smooth muscle cells.

受神经支配的、具有生理功能的平滑肌结构中的肠神经分化受到括约肌平滑肌细胞分泌的骨形态发生蛋白2的调节。

DOI：
10.1002/term.2027
发表时间：
2017
期刊：
Journal of tissue engineering and regenerative medicine
影响因子：
3.3
作者：
Rego,StephenL;Raghavan,Shreya;Zakhem,Elie;Bitar,KhalilN
通讯作者：
Bitar,KhalilN

Regenerative medicine as applied to general surgery.

再生医学应用于普通外科。

DOI：
10.1097/sla.0b013e318243a4db
发表时间：
2012-05
期刊：
Annals of surgery
影响因子：
9
作者：
Orlando G;Wood KJ;De Coppi P;Baptista PM;Binder KW;Bitar KN;Breuer C;Burnett L;Christ G;Farney A;Figliuzzi M;Holmes JH 4th;Koch K;Macchiarini P;Mirmalek Sani SH;Opara E;Remuzzi A;Rogers J;Saul JM;Seliktar D;Shapira-Schweitzer K;Smith T;Solomon D;Van Dyke M;Yoo JJ;Zhang Y;Atala A;Stratta RJ;Soker S
通讯作者：
Soker S

Design strategies of biodegradable scaffolds for tissue regeneration.