Rectoanal incontinence (RI) is one of the most debilitating gastrointestinal motility disorders affecting the elderly. A decrease in tone or stiffess of the smooth muscle of the internal anal sphincter (IAS) occurs with age and contributes to RI. Studies to date from our lab have provided insight into the regulatory control of IAS. In this competitive renewal application we propose to delineate novel control mechanisms of IAS muscle tone that can be targeted therapeutically. We will test the central hypothesis that miRNA regulation of RhoA/ROCK and SM22 is mechanistically linked to the age-dependent decrease in IAS tone and to RI, and that therapeutic targeting of miRNAs can increase IAS tone in vivo. We will establish differences in the signaling and contractile properties among multiple smooth muscle types of the lower GI tract;and in IAS as a function of age. Using both human and rat IAS, we propose studies to (Aim: 1) identify those intracellular signaling molecules that regulate IAS tone, and (Aim 2) identify miRNAs that regulate these intracellular signaling molecules, and develop strategies to reverse the miRNA-regulated decrease in IAS tone with age. We will establish multiple layers of control, focusing on the role of miRNAs as critical regulators of RhoA, RhoA-associated kinase (ROCK) and SM22. Preliminary biochemical and functional data support our hypothesis asserting the roles of RhoA, ROCK, and SM22 in determining SM tone among different SM types and age groups, while miRNA microarray analyses identify miR-139-5p and miR-1 as important regulators of ROCK and SM22 expression, respectively. Our robust approach includes cell, tissue, and in vivo models, enabling coordinated analysis of the miRNA-dependent proteome and signaling changes with functional effects. Beyond the unquestionable significance of the theme, the strengths of this proposal include: 1) exploration of novel mechanisms that explain the decrease in the IAS tone with age;2) a comprehensive and coordinated "omic", signaling and functional analysis that spans the reductionist- integrative spectrum;and 3) the development of a feasible therapeutic strategy for a disease that is currently untreatable.

Public Health Relevance

Rectoanal incontinence (RI), one of the most debilitating gastrointestinal motility disorders affecting the elderly, is characterized with decrease in tone o stiffness of the smooth muscle of the internal anal sphincter (IAS). Using model systems in the animals and humans, we will identify intracellular signaling molecules and novel regulatory microRNAs in the pathophysiology and therapeutic targeting for RI in the elderly.

Agency
National Institute of Health (NIH)
Type
Research Project (R01)
Project #
2R01DK035385-31A1
Application #
8760846
Study Section
Clinical, Integrative and Molecular Gastroenterology Study Section (CIMG)
Program Officer
Hamilton, Frank A
Project Start
Project End
Budget Start
Budget End
Support Year
31
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Thomas Jefferson University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
City
Philadelphia
State
PA
Country
United States
Zip Code
19107
Krishna, Chadalavada Vijay; Singh, Jagmohan; Kumar, Sumit et al. (2014) Heme oxygenase-1 upregulation modulates tone and fibroelastic properties of internal anal sphincter. Am J Physiol Gastrointest Liver Physiol 307:G595-601
Singh, Jagmohan; Kumar, Sumit; Krishna, Chadalavada Vijay et al. (2014) Aging-associated oxidative stress leads to decrease in IAS tone via RhoA/ROCK downregulation. Am J Physiol Gastrointest Liver Physiol 306:G983-91
Rattan, Satish (2013) Smooth muscle-specific myosin phosphatase target subunit 1 (MYPT1): an important piece of the puzzle. Gastroenterology 145:1494-5
Gibbons, Ahmara V; Lin, Jieru E; Kim, Gilbert W et al. (2013) Intestinal GUCY2C prevents TGF-* secretion coordinating desmoplasia and hyperproliferation in colorectal cancer. Cancer Res 73:6654-66
Romer, Anthony I; Singh, Jagmohan; Rattan, Satish et al. (2013) Smooth muscle fascicular reorientation is required for esophageal morphogenesis and dependent on Cdo. J Cell Biol 201:309-23
Singh, Jagmohan; Rattan, Satish (2013) Role of PKC and RhoA/ROCK pathways in the spontaneous phasic activity in the rectal smooth muscle. Am J Physiol Gastrointest Liver Physiol 304:G723-31
Rattan, Satish; Singh, Jagmohan (2012) RhoA/ROCK pathway is the major molecular determinant of basal tone in intact human internal anal sphincter. Am J Physiol Gastrointest Liver Physiol 302:G664-75
Singh, Jagmohan; Maxwell 4th, Pinckney J; Rattan, Satish (2011) Immunocytochemical evidence for PDBu-induced activation of RhoA/ROCK in human internal anal sphincter smooth muscle cells. Am J Physiol Gastrointest Liver Physiol 301:G317-25
de Godoy, Marcio A F; Rattan, Satish (2011) Role of rho kinase in the functional and dysfunctional tonic smooth muscles. Trends Pharmacol Sci 32:384-93
Rattan, S; Singh, J (2011) Basal internal anal sphincter tone, inhibitory neurotransmission, and other factors contributing to the maintenance of high pressures in the anal canal. Neurogastroenterol Motil 23:3-7

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