ADAM12: a genetic modifier of preclinical peripheral arterial disease

Dokun, Ayotunde O.; Chen, Lingdan; Okutsu, Mitsuharu; Farber, Charles R.; Hazarika, Surovi; Jones, W. Schuyler; Craig, Damian M.; Marchuk, Douglas A.; Lye, R. John; Shah, Svati H.; Annex, Brian H.

doi:10.1152/ajpheart.00803.2014

Cited by 34 publications

(97 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…We then used unilateral femoral artery ligation and resection (hind limb ischemia, HLI) in Balb/c mice, as a pre-clinical experimental model for severe PAD (critical limb ischemia, CLI-PAD) (22,23,24,25,26) . To determine whether VEGF 165 b induction correlates with decreased activation of VEGFR1 vs. VEGFR2 in ischemic muscle compared to non-ischemic we analyzed cells from non-ischemic and ischemic whole muscle tissue by flow cytometry (see Online Figure II for VEGF 165 b, pVEGFR1 Y1333 /VEGFR1, pVEGFR2 Y1175 /VEGFR2.…”

Section: Resultsmentioning

confidence: 99%

VEGF ₁₆₅ b Modulates Endothelial VEGFR1–STAT3 Signaling Pathway and Angiogenesis in Human and Experimental Peripheral Arterial Disease

Ganta

Choi

Kutateladze

et al. 2017

Circulation Research

Self Cite

118

View full text Add to dashboard Cite

Rationale Atherosclerotic-arterial occlusions decrease tissue perfusion causing ischemia to lower limbs in patients with peripheral arterial disease (PAD). Ischemia in muscle induces an angiogenic response but the magnitude of this response is frequently inadequate to meet tissue perfusion requirements. Alternate splicing in the exon-8 of vascular endothelial growth factor (VEGF)-A results in production of pro-angiogenic VEGFxxxa isoforms (VEGF165a, 165 for the 165 amino acid product) and anti-angiogenic VEGFxxxb (VEGF165b) isoforms. Objective The anti-angiogenic VEGFxxxb isoforms are thought to antagonize VEGFxxxa isoforms and decrease activation of VEGF-Receptor-2 (VEGFR2), hereunto considered the dominant receptor in post-natal angiogenesis in PAD. Our data will show that VEGF165b inhibits VEGFR1-Signal Transducer and Activator of Transcription (STAT)-3 signaling to decrease angiogenesis in human and experimental PAD. Methods and Results In human PAD vs. control muscle-biopsies, VEGF165b: a) is elevated, b) is bound higher (vs. VEGF165a) to VEGFR1 not VEGFR2, and c) levels correlated with decreased VEGFR1, not VEGFR2, activation. In experimental PAD, delivery of an isoform specific monoclonal antibody (Ab) to VEGF165b vs. control-Ab enhanced perfusion in animal model of severe PAD (Balb/c strain) without activating VEGFR2-signaling but with increased VEGFR1-activation. Receptor pull-down experiments demonstrate that VEGF165b-inhibition vs. control increased VEGFR1-STAT3 binding and STAT3-activation, independent of janus activated kinase (Jak1)/Jak2. Using VEGFR1+/− mice that could not increase VEGFR1 after ischemia, we confirm that VEGF165b decreases VEGFR1-STAT3 signaling to decrease perfusion. Conclusions Our results indicate that VEGF165b prevents activation of VEGFR1-STAT3 signaling by VEGF165a and hence inhibits angiogenesis and perfusion recovery in PAD muscle.

show abstract

Section: Resultsmentioning

confidence: 99%

VEGF ₁₆₅ b Modulates Endothelial VEGFR1–STAT3 Signaling Pathway and Angiogenesis in Human and Experimental Peripheral Arterial Disease

Ganta

Choi

Kutateladze

et al. 2017

Circulation Research

Self Cite

118

View full text Add to dashboard Cite

show abstract

“…This suggests the possible role of genetics in modifying the clinical outcome of an ischemic limb once an arterial occlusion is present. This is supported by preclinical animal studies where the recovery from the same degree of arterial injury is different between different genetic strains (69, 70), and several genes modify the clinical outcome from the same degree of lower extremity arterial injury (71–73). …”

Section: Pad and Geneticsmentioning

confidence: 90%

“…This region contains 495 genes. Further studies from our group have linked genes in the limb salvage quantitative trait locus 1 region to differences in PAD severity (71, 72). Wang et al showed that interleukin-21 receptor (IL-21R) is upregulated in endothelial cells from ischemic hind-limb muscle in mice and that loss of IL-21R resulted in impaired perfusion recovery (72).…”

Section: Future Of Genetic Studies In Padmentioning

confidence: 99%

Biomarkers and Genetics in Peripheral Artery Disease

Hazarika

Annex

2017

Clinical Chemistry

Self Cite

View full text Add to dashboard Cite

BACKGROUND Peripheral artery disease (PAD) is highly prevalent and there is considerable diversity in the initial clinical manifestation and disease progression among individuals. Currently, there is no ideal biomarker to screen for PAD, to risk stratify patients with PAD, or to monitor therapeutic response to revascularization procedures. Advances in human genetics have markedly enhanced the ability to develop novel diagnostic and therapeutic approaches across a host of human diseases, but such developments in the field of PAD are lagging. CONTENT In this article, we will discuss the epidemiology, traditional risk factors for, and clinical presentations of PAD. We will discuss the possible role of genetic factors and gene–environment interactions in the development and/or progression of PAD. We will further explore future avenues through which genetic advances can be used to better our understanding of the pathophysiology of PAD and potentially find newer therapeutic targets. We will discuss the potential role of biomarkers in identifying patients at risk for PAD and for risk stratifying patients with PAD, and novel approaches to identification of reliable biomarkers in PAD. SUMMARY The exponential growth of genetic tools and newer technologies provides opportunities to investigate and identify newer pathways in the development and progression of PAD, and thereby in the identification of newer biomarkers and therapies.

show abstract

“…In vitro, augmentation of ADAM12 expression improved endothelial cell (EC) proliferation and angiogenesis in ischemia while knocking down ADAM12 impaired proliferation, survival, and angiogenesis in ischemia. 12 These findings suggest that ADAM12 plays an important role in post-ischemic angiogenesis but less is known about the mechanisms regulating ADAM12 expression in ischemia.…”

Section: Introductionmentioning

confidence: 99%

Modulation of miR29a improves impaired post-ischemic angiogenesis in hyperglycemia

Chen

Okeke

Ayalew

et al. 2017

Exp Biol Med (Maywood)

Self Cite

View full text Add to dashboard Cite

Individuals with diabetes mellitus suffer from impaired angiogenesis and this contributes to poorer peripheral arterial disease outcomes. In experimental peripheral arterial disease, angiogenesis and perfusion recovery are impaired in mice with diabetes. We recently showed that a disintegrin and metalloproteinase domain-containing protein 12 (ADAM12) is upregulated in ischemic endothelial cells and plays a key role in post-ischemic angiogenesis and perfusion recovery following experimental peripheral arterial disease. Here we investigated the role of miR29a in the regulation of endothelial cell ADAM12 expression in ischemia and how hyperglycemia negatively affects this regulation. We also explored whether modulating miR29a can improve impaired post-ischemic angiogenesis associated with hyperglycemia. Additionally, we tested whether miR29a modulation could improve post ischemic angiogenesis in the setting of impaired vascular endothelial growth factor signaling. We forced miR29a expression in ischemic endothelial cells and assessed ADAM12 expression. We also evaluated whether hyperglycemia in vivo and in vitro impair ischemia-induced ADAM12 upregulation and miR29a downregulation. Lastly, we determined whether modulating endothelial cell miR29a expression in ischemia and hyperglycemia could improve impaired endothelial cell functions. We found under ischemic conditions where ADAM12 is upregulated in endothelial cells, miR29a is downregulated. Forced expression of miR29a in ischemic endothelial cell prevented ADAM12 upregulation . In ischemic hind limbs of mice with type 1 diabetes and in endothelial cells exposed to simulated ischemia plus hyperglycemia, ADAM12 upregulation and miR29a downregulation were blunted while angiogenesis was impaired. Knocking down miR29a with an miR29a inhibitor was sufficient to improve ADAM12 upregulation and angiogenesis in simulated ischemia plus hyperglycemia. It was also sufficient to improve perfusion recovery in type 1 diabetes mellitus mice in vivo and angiogenesis in vitro even when vascular endothelial growth factor signaling was impaired with blocking antibodies. In conclusion, MiR29a regulates endothelial cell ADAM12 upregulation in ischemia and this is impaired in hyperglycemia. Modulating miR29a improves impaired post-ischemic angiogenesis associated with hyperglycemia. Impact statement Individuals with diabetes are more likely to develop peripheral arterial disease (PAD), and when PAD is present, in those with diabetes, it is more severe and there is currently no effective medical treatment for impaired blood flow which occurs in diabetics with PAD. The current work advances the field by providing an understanding of a molecular mechanism involved in impaired post ischemic angiogenesis in diabetes. It shows for the first time that failure to downregulate miR29a in ischemic diabetic tissues is a major contributing factor to poor perfusion recovery in experimental PAD, and miR29a is elevated in skeletal muscle samples from human diabetics compared with levels in those without diabetes. Knocking down the elevated miR29a in ischemic diabetic mouse hind limbs improved perfusion recovery following experimental PAD. This shows miR29a modulation as a novel therapeutic target for improving blood flow in diabetics with PAD.

show abstract

ADAM12: a genetic modifier of preclinical peripheral arterial disease

Cited by 34 publications

References 37 publications

VEGF ₁₆₅ b Modulates Endothelial VEGFR1–STAT3 Signaling Pathway and Angiogenesis in Human and Experimental Peripheral Arterial Disease

VEGF ₁₆₅ b Modulates Endothelial VEGFR1–STAT3 Signaling Pathway and Angiogenesis in Human and Experimental Peripheral Arterial Disease

Biomarkers and Genetics in Peripheral Artery Disease

Modulation of miR29a improves impaired post-ischemic angiogenesis in hyperglycemia

Contact Info

Product

Resources

About

ADAM12: a genetic modifier of preclinical peripheral arterial disease

Cited by 34 publications

References 37 publications

VEGF 165 b Modulates Endothelial VEGFR1–STAT3 Signaling Pathway and Angiogenesis in Human and Experimental Peripheral Arterial Disease

VEGF 165 b Modulates Endothelial VEGFR1–STAT3 Signaling Pathway and Angiogenesis in Human and Experimental Peripheral Arterial Disease

Biomarkers and Genetics in Peripheral Artery Disease

Modulation of miR29a improves impaired post-ischemic angiogenesis in hyperglycemia

Contact Info

Product

Resources

About

VEGF ₁₆₅ b Modulates Endothelial VEGFR1–STAT3 Signaling Pathway and Angiogenesis in Human and Experimental Peripheral Arterial Disease

VEGF ₁₆₅ b Modulates Endothelial VEGFR1–STAT3 Signaling Pathway and Angiogenesis in Human and Experimental Peripheral Arterial Disease