Joseph Miano

Joseph Miano
Medical College of Georgia at Augusta University?·?Vascular Biology

54.76
?·?
PhD

About

176
Publications
68,495
Reads
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Introduction
My lab 'mines' the human genome for functional noncoding sequences such as regulatory elements and long non-coding RNA genes to gain insight into the control of vascular smooth muscle cell differentiation and to understand regulatory variants in the CArG box (binds SRF) associated with human diseases. My lab uses computers, human cell culture models and genetically tractable mouse models, especially those generated using the revolutionary CRISPR-Cas9 system of genome editing.
Research Experience
November 2019 - present
Medical College of Georgia at Augusta University
Position
  • J Harold Harrison Distinguished University Chair in Vascular Biology
October 2015 - present
University of Rochester
Position
  • Professor
July 2003 - October 2015
University of Rochester
Position
  • Associate Professor

Publications

Publications (176)
Article
Full-text available
To ascertain the importance of a single regulatory element in the control of Cnn1 expression using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 genome editing. The CRISPR/CRISPR-associated protein 9 system was used to produce 3 of 18 founder mice carrying point mutations in an intronic CArG box of t...
Article
Full-text available
Previous efforts to target the mouse genome for the addition, subtraction, or substitution of biologically informative sequences required complex vector design and a series of arduous steps only a handful of laboratories could master. The facile and inexpensive clustered regularly interspaced short palindromic repeats (CRISPR) method has now supers...
Article
Full-text available
Megacystis microcolon intestinal hypoperistalsis syndrome (MMIHS) is a congenital visceral myopathy characterized by severe dilation of the urinary bladder and defective intestinal motility. The genetic basis of MMIHS has been ascribed to spontaneous and autosomal dominant mutations in actin gamma 2 (ACTG2), a smooth muscle contractile gene. Howeve...
Experiment Findings
Full-text available
Article
Epidemiological studies suggest that individuals in the Mediterranean region with a loss-of-function, nonsynonymous single nucleotide polymorphism (S188F), in glucose-6-phosphate dehydrogenase (G6pd) are less susceptible to vascular diseases. However, this association has not yet been experimentally proven. Here, we set out to determine whether the...
Article
Full-text available
Rationale: The gene encoding TCF21 (transcription factor 21) has been linked to coronary artery disease risk by human genome-wide association studies in multiple racial ethnic groups. In murine models, Tcf21 is required for phenotypic modulation of smooth muscle cells (SMCs) in atherosclerotic tissues and promotes a fibroblast phenotype in these c...
Article
Full-text available
SENCR is a human-specific, vascular cell-enriched long-noncoding RNA (lncRNA) that regulates vascular smooth muscle cell and endothelial cell (EC) phenotypes. The underlying mechanisms of action of SENCR in these and other cell types is unknown. Here, levels of SENCR RNA are shown to be elevated in several differentiated human EC lineages subjected...
Preprint
Full-text available
CRISPR–Cas9 gene editing technology has considerably facilitated the generation of mouse knockout alleles, relieving many of the cumbersome and time-consuming steps of traditional mouse embryonic stem cell technology. However, the generation of conditional knockout alleles remains an important challenge. An earlier study reported up to 16% efficien...
Article
Full-text available
Objective: Unreliable antibodies often hinder the accurate detection of an endogenous protein, and this is particularly true for the cardiac and smooth muscle cofactor, MYOCD (myocardin). Accordingly, the mouse Myocd locus was targeted with 2 independent epitope tags for the unambiguous expression, localization, and activity of MYOCD protein. App...
Article
Full-text available
Podocytes contain an intricate actin cytoskeleton that is essential for the specialized function of this cell type in renal filtration. Serum response factor (SRF) is a master transcription factor for the actin cytoskeleton, but the in vivo expression and function of SRF in podocytes are unknown. We found that SRF protein colocalizes with podocyte...
Article
Full-text available
Serum response factor (SRF) transcriptionally regulates expression of contractile genes in smooth muscle cells (SMC). Lack or decrease of SRF is directly linked to a phenotypic change of SMC, leading to hypomotility of smooth muscle in the gastrointestinal (GI) tract. However, the molecular mechanism behind SRF-induced hypomotility in GI smooth mus...
Data
Ca2+ transients and tissue movement activities recorded from longitudinal smooth muscle cells and ICC-MY in Srf WT jejunum within a field of view. *QuickTime Player is required. (MOV)
Data
Oligonucleotides used in this study. (DOCX)
Data
A list of proteins that were down-regulated in the jejunal smooth muscle of Srf KO mice. (XLSX)
Data
Analysis of SRF binding sites and CArG boxes of SRF down-regulated protein genes. (XLS)
Article
Full-text available
Objective: To identify and characterize the effect of a SNP (single-nucleotide polymorphism) in the STXBP5 locus that is associated with altered thrombosis in humans. GWASs (genome-wide association studies) have identified numerous SNPs associated with human thrombotic phenotypes, but determining the functional significance of an individual candid...
Article
Early parental loss is associated with social–emotional dysregulation and amygdala physiologic changes. Previously, we examined whole amygdala gene expression in infant monkeys exposed to early maternal deprivation. Here, we focus on an amygdala region with immature neurons at birth: the paralaminar nucleus (PL). We hypothesized that 1) the normal...
Article
The new millennium heralds an unanticipated surge of genomic information, most notably an expansive class of long noncoding RNAs (lncRNAs). These transcripts, which now outnumber all protein-coding genes, often exhibit the same characteristics as mRNAs (RNA polymerase II-dependent, 5' methyl-capped, multiexonic, polyadenylated); yet, they do not en...
Article
Prostatic smooth muscle cells (pSMCs) differentiation is a key factor for prostatic 顺心彩票ostasis, with androgens exerting multiple effects on these cells. Here, we demonstrated that the myodifferentiator complex Srf/Myocd is up-regulated by testosterone in a dose-dependent manner in primary cultures of rat pSMCs, which was associated to the increase...
Article
Full-text available
Homeostatic control of vascular smooth muscle cell (VSMC) differentiation is critical for contractile activity and regulation of blood flow. Recently, we reported that pre-contracted blood vessels are relaxed and the phenotype of VSMC is regulated from a synthetic to contractile state by glucose-6-phosphate dehydrogenase (G6PD) inhibition. In the c...
Article
Objective: Long noncoding RNAs (lncRNA) represent a growing class of noncoding genes with diverse cellular functions. We previously reported on SENCR, an lncRNA that seems to support the vascular smooth muscle cell (VSMC) contractile phenotype. However, information about the VSMC-specific lncRNAs regulated by myocardin (MYOCD)/serum response facto...
Article
Full-text available
Background: -Phenotypic switching of vascular smooth muscle cells (VSMCs) from a contractile to a synthetic state is implicated in diverse vascular pathologies including atherogenesis, plaque stabilisation, and neointimal hyperplasia. However, very little is known as to the role of long non coding RNA (lncRNA) during this process. Here we investig...
Article
Full-text available
Despite the increasing importance of long non-coding RNA in physiology and disease, their role in endothelial biology remains poorly understood. Growing evidence has highlighted them to be essential regulators of human embryonic stem cell differentiation. SENCR, a vascular-enriched long non-coding RNA, overlaps the Friend Leukemia Integration virus...
Article
Full-text available
Serum response factor (SRF) is a transcription factor known to mediate phenotypic plasticity in smooth muscle cells (SMCs). Despite the critical role of this protein in mediating intestinal injury response, little is known about the mechanism through which SRF alters SMC behavior. Here, we provide compelling evidence for the involvement of SRF-depe...
Article
Full-text available
Background/aims: Smooth muscle cells (SMCs) characteristically express serum response factor (SRF), which regulates their development. The role of SRF in SMC plasticity in the pathophysiological conditions of gastrointestinal (GI) tract is less characterized. Methods: We generated SMC-specific Srf knockout mice and characterized the prenatally l...
Article
Full-text available
Caveolae are membrane organelles that play roles in glucose and lipid metabolism and in vascular function. Formation of caveolae requires caveolins and cavins. The make-up of caveolae and their density is considered to reflect cell-specific transcriptional control mechanisms for caveolins and cavins, but knowledge regarding regulation of caveolae g...
Article
Full-text available
Genome-scale expression data on the absolute numbers of gene isoforms offers essential clues in cellular functions and biological processes. Smooth muscle cells (SMCs) perform a unique contractile function through expression of specific genes controlled by serum response factor (SRF), a transcription factor that binds to DNA sites known as the CArG...
Article
Full-text available
The two principal cell types of importance for normal vessel wall physiology are smooth muscle cells and endothelial cells. Much progress has been made over the past 20 years in the discovery and function of transcription factors that coordinate proper differentiation of these cells and the maintenance of vascular 顺心彩票ostasis. More recently, the co...
Article
Full-text available
Myocardin (MYOCD) is a potent transcriptional coactivator that functions primarily in cardiac muscle and smooth muscle through direct contacts with serum response factor (SRF) over cis elements known as CArG boxes found near a number of genes encoding for contractile, ion channel, cytoskeletal, and calcium handling proteins. Since its discovery mor...
Article
Atherosclerosis, the cause of 50% of deaths in westernized societies, is widely regarded as a chronic vascular inflammatory disease. Vascular smooth muscle cell (VSMC) inflammatory activation in response to local proinflammatory stimuli contributes to disease progression and is a pervasive feature in developing atherosclerotic plaques. Therefore, i...
Article
We previously showed that cholesterol loading in vitro converts mouse aortic vascular smooth muscle cells (VSMC) from a contractile state to one resembling macrophages. In human and mouse atherosclerotic plaques, it has become appreciated that ≈40% of cells classified as macrophages by histological markers may be of VSMC origin. Therefore, we sough...
Article
Full-text available
Smooth muscle cells are maintained in a differentiated state in the vessel wall, but can be modulated to a synthetic phenotype following injury. Smooth muscle phenotypic modulation is thought to play an important role in the pathology of vascular occlusive diseases. Phenotypically modulated smooth muscle cells exhibit increased proliferative and mi...
Article
Vascular smooth muscle cells (VSMCs) play important physiological and pathophysiological roles. Two articles in the current issue focus on the latter in atherosclerosis1 and in pulmonary arterial hypertension (PAH).2 Notably, Allahverdian et al1 report that, in human coronary artery atherosclerotic plaques, many cells typically classified as macrop...
Article
Long noncoding RNAs (lncRNAs) represent a rapidly growing class of RNA genes with functions related primarily to transcriptional and post-transcriptional control of gene expression. There is a paucity of information about lncRNA expression and function in human vascular cells. Thus, we set out to identify novel lncRNA genes in human vascular smooth...
Data
##Assembly-Data-START## Assembly Method :: SHRiMP v. 2.2.3 Sequencing Technology :: Illumina; Sanger dideoxy sequencing ##Assembly-Data-END##
Article
Overactive bladder (OAB) is a pervasive clinical problem involving alterations in both neurogenic and myogenic activity. While there has been some progress in understanding neurogenic inputs to OAB, the mechanisms controlling myogenic bladder activity are unclear. We report the involvement of myocardin (MYOCD) and microRNA-1 (miR-1) in the regulati...
Article
Full-text available
Activated platelets release many inflammatory molecules with important roles in accelerating vascular inflammation. Much is known about platelet and platelet-derived mediator interactions with endothelial cells and leukocytes, but few studies have examined the effects of platelets on components of the vascular wall. Vascular smooth muscle cells (VS...
Article
Vascular smooth muscle cell (VSMC) differentiation is determined primarily by the level and activity of serum response factor and myocardin, which constitute a potent transcriptional switch over CArG elements located near a growing number of VSMC protein coding and non-coding RNA genes. Unlike the other two muscle types, differentiated VSMC are int...
Article
Objective: Several studies have shown through chemical inhibitors that p38 mitogen-activated protein kinase (MAPK) promotes vascular smooth muscle cell (VSMC) differentiation. Here, we evaluate the effects of knocking down a dominant p38MAPK isoform on VSMC differentiation. Methods and results: Knockdown of p38MAPKα (MAPK14) in human coronary ar...
Article
Full genome annotation requires gene expression analysis and elucidation of promoter activity. Here, we analyzed the expression and promoter of a highly restricted integrin gene, Itga8. RNase protection and quantitative RT-PCR showed Itga8 to be expressed most abundantly in vascular smooth muscle cells (SMC). Transcription start site mapping of Itg...
Article
More than 50 years ago, smooth muscle cells (SMC) of the carotid artery were shown to undergo "de-differentiation" upon ligation injury.(1) Since this classic study, scores of research groups have used a variety of in vivo and in vitro model systems as well as numerous clinical studies to demonstrate the conversion of normally contractile vascular...
Article
Benign prostatic hyperplasia (BPH) and bladder outlet obstruction (BOO) are common in older men and can contribute to lower urinary tract symptoms that significantly impact quality of life. Few existing models of BOO and BPH use physiological levels of hormones associated with disease progression in humans in a genetically manipulable organism. We...
Article
Full-text available
The advent of modern mouse genetics has benefited many fields of diseased-based research over the past 20 years, none perhaps more profoundly than cardiac biology. Indeed, the heart is now arguably one of the easiest tissues to genetically manipulate, given the availability of an ever-growing tool chest of molecular reagents/promoters and "facilita...
Article
Serum response factor (SRF) plays vital roles in numerous cellular processes; however, the physiological function of SRF in skeletal tissue remains unknown. In several organ systems, SRF regulates the expression of insulin-like growth factor-1 (IGF-1), which is crucial for normal development of mineralized skeleton and bone remodeling throughout li...
Article
Full-text available
Smooth muscle cell (SMC) differentiation is defined largely by a number of cell-restricted genes governed directly by the serum response factor (SRF)/myocardin (MYOCD) transcriptional switch. Here, we describe a new SRF/MYOCD-dependent, SMC-restricted gene known as Leiomodin 1 (Lmod1). Conventional and quantitative RT-PCRs indicate that Lmod1 mRNA...
Article
See related article, pages 880–893 The human genome is replete with digital information, only 1.2% of which comprises protein-coding sequence. The nonprotein-coding sequence encompasses >1 million regulatory elements controlling gene expression and codes for such genomic processes as recombination, replication, splicing, transposition, and structu...
Article
Smooth muscle calponin (CNN1) contains multiple conserved intronic CArG elements that bind serum response factor and display enhancer activity in vitro. The objectives here were to evaluate these CArG elements for activity in transgenic mice and determine the effect of human CNN1 on injury-induced vascular remodeling. Mice carrying a lacZ reporter...
Article
Full-text available
Regulatory SNPs (rSNPs) reside primarily within the nonprotein coding genome and are thought to disturb normal patterns of gene expression by altering DNA binding of transcription factors. Nevertheless, despite the explosive rise in SNP association studies, there is little information as to the function of rSNPs in human disease. Serum response fac...
Article
Full-text available
microRNA (miR) 143/145 is restricted to adult smooth muscle cell (SMC) lineages and mediates, in part, the expression of several SMC contractile genes. Although the function of miR143/145 has begun to be elucidated, its transcriptional regulation in response to various signaling inputs is poorly understood. In an effort to define a miR signature fo...
Article
Full-text available
MicroRNA 143/145 (miR143/145) is restricted to adult smooth muscle cell (SMC) lineages and mediates, in part, the expression of several SMC contractile genes. Although the function of miR143/145 has begun to be elucidated, its transcriptional regulation in response to various signaling inputs is poorly understood. In an effort to define a miR signa...
Article
Full-text available
Both TGF-β and myocardin (MYOCD) are important for smooth muscle cell (SMC) differentiation, but their precise role in regulating the initiation of SMC development is less clear. In TGF-β-induced SMC differentiation of pluripotent C3H10T1/2 progenitors, we found that TGF-β did not significantly induce Myocd mRNA expression until 18 h of stimulation...
Data
RT-qPCR analysis of selected mRNAs in the aorta of Cre-negative (Cre-) mice, 10 weeks post vehicle (Veh) or Tamoxifen (Tx) treatment. (TIFF)
Data
Tail DNA from Cre recombinase positive mice, wild type, homozygous or heterozygous for the floxed Dicer allele were genotyped before and two weeks after tamoxifen treatment. A representative agarose gel, which displays the presence of band representing the excised gene after tamoxifen treatment is shown in A. (B) analysis of body weight and tibia l...
Data
Transmission electron micrographs of aortic SMCs of control and Dicer KO mice, 10 weeks post tamoxifen tratament. (A) Aortic SMC from control mouse. Note typical abundance of myofilaments in the cytosol of two adjacent medial SMC (black arrows) as well as peripheral dense plaques (arrowheads). (B) Aortic SMC from SM-Dicer KO mouse. Note the virtual...
Data
(A) Relative serotonin (5HT)-induced calcium influx in isolated Ctrl and KO isolated aortic SMCs were measured using Fluo-4 calcium indicator (A). (B-I) Contractile function of control and SM Dicer KO arteries 5 weeks post Tamoxifen treatment. (B) Active force of small mesenteric arteries in response to 80 mM KCl (HK). (C) Summarized data of the pa...
Data
(A-E) Morphological analysis and analysis of cell number of the aorta of Cre-negative (Cre-) mice, 10 weeks post Tamoxifen (Tx) or vehicle (Veh) treatment. (F-J) Perfusion fixed and paraffin embedded sections of the saphenous artery of control (Ctrl) and SM-Dicer KO (KO) mice were analyzed for morphological changes and cell number. (K) Representati...
Article
Full-text available
Phenotypic modulation of smooth muscle cells (SMCs) plays a key role in vascular disease, including atherosclerosis. Several transcription factors have been suggested to regulate phenotypic modulation of SMCs but the decisive mechanisms remain unknown. Recent reports suggest that specific microRNAs (miRNAs) are involved in SMC differentiation and v...
Data
Non-immunogenic IgG control antisera was applied to uninjured right carotid (A), 7 day injured left carotid (B), and 21 day injured left carotid (C). Panels D-F represent AKAP12 staining of uninjured carotid (D) and femoral (E) artery or a 7 day complete ligation injured carotid artery (F). Note loss of AKAP12 staining in the media of the injured v...