microRNA‐29a Regulates ADAM12 Through Direct Interaction With ADAM12 mRNA and Modulates Postischemic Perfusion Recovery
Background Peripheral artery disease is caused by atherosclerotic occlusion of vessels outside the heart and most commonly affects vessels of the lower extremities. Angiogenesis is a part of the postischemic adaptation involved in restoring blood flow in peripheral artery disease. Previously, in a m...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2022-08-01
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| Series: | Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease |
| Subjects: | |
| Online Access: | https://www.ahajournals.org/doi/10.1161/JAHA.122.025727 |
| _version_ | 1811215175284424704 |
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| author | Victor Lamin Joseph Verry Olumayowa S. Dokun Ana Kronemberger Thomas Wong Vitor A. Lira Ayotunde O. Dokun |
| author_facet | Victor Lamin Joseph Verry Olumayowa S. Dokun Ana Kronemberger Thomas Wong Vitor A. Lira Ayotunde O. Dokun |
| author_sort | Victor Lamin |
| collection | DOAJ |
| description | Background Peripheral artery disease is caused by atherosclerotic occlusion of vessels outside the heart and most commonly affects vessels of the lower extremities. Angiogenesis is a part of the postischemic adaptation involved in restoring blood flow in peripheral artery disease. Previously, in a murine hind limb ischemia model of peripheral artery disease, we identified ADAM12 (a disintegrin and metalloproteinase gene 12) as a key genetic modifier of postischemic perfusion recovery. However, less is known about ADAM12 regulation in ischemia. MicroRNAs are a class of small, noncoding, single‐stranded RNAs that regulate gene expression primarily through transcriptional repression of messenger RNA (mRNA). We showed microRNA‐29a (miR‐29a) modulates ADAM12 expression in the setting of diabetes and ischemia. However, how miR‐29a modulates ADAM12 is not known. Moreover, the physiological effects of miR‐29a modulation in a nondiabetic setting is not known. Methods and Results We overexpressed or inhibited miR‐29a in ischemic mouse gastrocnemius and tibialis anterior muscles, and quantified the effect on perfusion recovery, ADAM12 expression, angiogenesis, and skeletal muscle regeneration. In addition, using RNA immunoprecipitation–based anti‐miR competitive assay, we investigated the interaction of miR‐29a and ADAM12 mRNA in mouse microvascular endothelial cell, skeletal muscle, and human endothelial cell lysates. Ectopic expression of miR‐29a in ischemic mouse hind limbs decreased ADAM12 mRNA expression, increased skeletal muscle injury, decreased skeletal muscle function, and decreased angiogenesis and perfusion recovery, with no effect on skeletal muscle regeneration and myofiber cross‐sectional area following hind limb ischemia. RNA immunoprecipitation–based anti‐miR competitive assay studies showed miR‐29a antagomir displaced miR‐29a and ADAM12 mRNA from the AGO‐2 (Argonaut‐2) complex in a dose dependent manner. Conclusions Taken together, the data show miR‐29a suppresses ADAM12 expression by directly binding to its mRNA, resulting in impaired skeletal muscle function, angiogenesis, and poor perfusion. Hence, elevated levels of miR‐29a, as seen in diabetes and aging, likely contribute to vascular pathology, and modulation of miR‐29a could be a therapeutic target. |
| first_indexed | 2024-04-12T06:17:35Z |
| format | Article |
| id | doaj.art-dc2f88d9f651400888d2476a78f60cc3 |
| institution | Directory Open Access Journal |
| issn | 2047-9980 |
| language | English |
| last_indexed | 2024-04-12T06:17:35Z |
| publishDate | 2022-08-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease |
| spelling | doaj.art-dc2f88d9f651400888d2476a78f60cc32022-12-22T03:44:25ZengWileyJournal of the American Heart Association: Cardiovascular and Cerebrovascular Disease2047-99802022-08-01111610.1161/JAHA.122.025727microRNA‐29a Regulates ADAM12 Through Direct Interaction With ADAM12 mRNA and Modulates Postischemic Perfusion RecoveryVictor Lamin0Joseph Verry1Olumayowa S. Dokun2Ana Kronemberger3Thomas Wong4Vitor A. Lira5Ayotunde O. Dokun6Division of Endocrinology and Metabolism, Carver College of Medicine University of Iowa Iowa City IADivision of Endocrinology and Metabolism, Carver College of Medicine University of Iowa Iowa City IADivision of Endocrinology and Metabolism, Carver College of Medicine University of Iowa Iowa City IAFraternal Order of Eagles Diabetes Research Center, Carver College of Medicine University of Iowa Iowa City IADivision of Endocrinology and Metabolism, Carver College of Medicine University of Iowa Iowa City IAFraternal Order of Eagles Diabetes Research Center, Carver College of Medicine University of Iowa Iowa City IADivision of Endocrinology and Metabolism, Carver College of Medicine University of Iowa Iowa City IABackground Peripheral artery disease is caused by atherosclerotic occlusion of vessels outside the heart and most commonly affects vessels of the lower extremities. Angiogenesis is a part of the postischemic adaptation involved in restoring blood flow in peripheral artery disease. Previously, in a murine hind limb ischemia model of peripheral artery disease, we identified ADAM12 (a disintegrin and metalloproteinase gene 12) as a key genetic modifier of postischemic perfusion recovery. However, less is known about ADAM12 regulation in ischemia. MicroRNAs are a class of small, noncoding, single‐stranded RNAs that regulate gene expression primarily through transcriptional repression of messenger RNA (mRNA). We showed microRNA‐29a (miR‐29a) modulates ADAM12 expression in the setting of diabetes and ischemia. However, how miR‐29a modulates ADAM12 is not known. Moreover, the physiological effects of miR‐29a modulation in a nondiabetic setting is not known. Methods and Results We overexpressed or inhibited miR‐29a in ischemic mouse gastrocnemius and tibialis anterior muscles, and quantified the effect on perfusion recovery, ADAM12 expression, angiogenesis, and skeletal muscle regeneration. In addition, using RNA immunoprecipitation–based anti‐miR competitive assay, we investigated the interaction of miR‐29a and ADAM12 mRNA in mouse microvascular endothelial cell, skeletal muscle, and human endothelial cell lysates. Ectopic expression of miR‐29a in ischemic mouse hind limbs decreased ADAM12 mRNA expression, increased skeletal muscle injury, decreased skeletal muscle function, and decreased angiogenesis and perfusion recovery, with no effect on skeletal muscle regeneration and myofiber cross‐sectional area following hind limb ischemia. RNA immunoprecipitation–based anti‐miR competitive assay studies showed miR‐29a antagomir displaced miR‐29a and ADAM12 mRNA from the AGO‐2 (Argonaut‐2) complex in a dose dependent manner. Conclusions Taken together, the data show miR‐29a suppresses ADAM12 expression by directly binding to its mRNA, resulting in impaired skeletal muscle function, angiogenesis, and poor perfusion. Hence, elevated levels of miR‐29a, as seen in diabetes and aging, likely contribute to vascular pathology, and modulation of miR‐29a could be a therapeutic target.https://www.ahajournals.org/doi/10.1161/JAHA.122.025727ADAM12angiogenesishind limb ischemiamiR‐29a and peripheral artery disease |
| spellingShingle | Victor Lamin Joseph Verry Olumayowa S. Dokun Ana Kronemberger Thomas Wong Vitor A. Lira Ayotunde O. Dokun microRNA‐29a Regulates ADAM12 Through Direct Interaction With ADAM12 mRNA and Modulates Postischemic Perfusion Recovery Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease ADAM12 angiogenesis hind limb ischemia miR‐29a and peripheral artery disease |
| title | microRNA‐29a Regulates ADAM12 Through Direct Interaction With ADAM12 mRNA and Modulates Postischemic Perfusion Recovery |
| title_full | microRNA‐29a Regulates ADAM12 Through Direct Interaction With ADAM12 mRNA and Modulates Postischemic Perfusion Recovery |
| title_fullStr | microRNA‐29a Regulates ADAM12 Through Direct Interaction With ADAM12 mRNA and Modulates Postischemic Perfusion Recovery |
| title_full_unstemmed | microRNA‐29a Regulates ADAM12 Through Direct Interaction With ADAM12 mRNA and Modulates Postischemic Perfusion Recovery |
| title_short | microRNA‐29a Regulates ADAM12 Through Direct Interaction With ADAM12 mRNA and Modulates Postischemic Perfusion Recovery |
| title_sort | microrna 29a regulates adam12 through direct interaction with adam12 mrna and modulates postischemic perfusion recovery |
| topic | ADAM12 angiogenesis hind limb ischemia miR‐29a and peripheral artery disease |
| url | https://www.ahajournals.org/doi/10.1161/JAHA.122.025727 |
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