Project Details
Description
PROJECT SUMMARY/ABSTRACT:
While portal hypertension accounts for significant mortality in patients with liver cirrhosis, its molecular
pathogenesis remains undefined. The Overall Objective of this grant is to define how intravascular pressures
increase in the hepatic sinusoid, which is the initiating step of portal hypertension. Liver injury leads to tissue
edema and early matrix deposition which increases stiffness in the interstitial Space of Disse. How mechanical
forces such as stiffness regulates endothelial cell signaling and gene transcription to influence portal pressure
is a gap in knowledge. Recent studies reveal unanticipated importance of glycolytic metabolism in diverse and
critical endothelial cell functions. In this regard, our Preliminary Data in liver endothelial cells (LEC) shows that
interstitial stiffness: 1) triggers glycolysis dependent remodeling of focal adhesions and actin stress fibers
which culminates in, 2) histone acetylation dependent production and release of the CXCL1 neutrophil
chemokine, and that 3) neutrophils can form neutrophil extracellular traps (NETS) and microthrombi in the
hepatic sinusoids. NETS are extruded nuclear proteins from neutrophils implicated in thrombosis. Indeed,
clinical evidence from patients with portal hypertension demonstrates microthrombi in the hepatic sinusoids
although their pathogenic role in portal hypertension has not been previously defined. We have utilized these
novel findings to generate the Central Hypothesis of the current proposal; that glycolysis dependent
mechanotransduction in LEC leads to CXCL1 release that mediates neutrophil derived sinusoidal microthrombi
to increase portal pressure. The Aims are to test the sub-hypotheses that: 1) Glycolysis is required for
mechanotransduction in LEC. Aim 1a will examine mechanisms how interstitial stiffness increases glycolytic
activity by recruiting glycolytic enzymes to focal adhesions for their activation. Aim 1b will test how glycolysis
induces actin polymerization to promote mechanotransduction from focal adhesions to the nucleus by
increasing nuclear pore size and enabling nuclear translocation of the mechanosensitive transcription activator
YAP. 2) LEC mechanotransduction leads to CXCL1 release that recruits neutrophils to LEC. Aim 2a will
explore how YAP recruits the histone acetyltransferase, p300, from the CXCL1 enhancer to the CXCL1
promoter to deposit histone marks that activate gene transcription. Aim 2b will examine functional effects of
released CXCL1 on neutrophil adhesion to LEC. 3) CXCL1 recruited neutrophils produce NETS leading to
sinusoidal microthrombi and portal hypertension. Aim 3 will use a combination of genetic and pharmacologic
interventions to disrupt glycolytic enzyme function and YAP-p300 activation of CXCL1 production and release,
to ascertain effects on portal hypertension development in mice in vivo. The Aim will also utilize innovative
imaging techniques including atomic force microscopy, magnetic resonance elastography, and intravital
microscopy to measure cellular and organ changes in stiffness in coordination with NETS, microthrombi, and
portal hypertension. Thus, this proposal will explore how mechanotransduction drives metabolism to regulate
chromatin and gene transcription in the hepatic sinusoids. This novel and innovative line of inquiry will define
an LEC dependent model of portal hypertension and set a trajectory towards new and significant advances to
treat portal hypertension in humans.
Status | Active |
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Effective start/end date | 5/15/02 → 5/31/24 |
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