Wnt/β-Catenin Stimulation and Laminins Support Cardiovascular Cell Progenitor Expansion from Human Fetal Cardiac Mesenchymal Stromal Cells

Månsson‐Broberg, Agneta; Rodin, Sergey; Bulatović, Ivana; Ibarra, Carmen; Löfling, Marie; Genead, Rami; Wärdell, Eva; Felldin, Ulrika; Granath, Carl; Alici, Evren; Blanc, Katarina Le; Smith, C. I. Edvard; Salašová, Alena; Westgren, Magnus; Sundström, Erik; Uhlén, Per; Arenas, Ernest; Sylvén, Christer; Tryggvason, Karl; Corbascio, Matthias; Simonson, Oscar E.; Österholm, Cecilia; Grinnemo, Karl‐Henrik

doi:10.1016/j.stemcr.2016.02.014

Cited by 21 publications

(21 citation statements)

References 54 publications

(96 reference statements)

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“…Mumtaz, et al showed that ZIKV infection perturbs osteoblast-derived MSC function without causing cytopathic effects, and additionally found a delay in osteoblast development and maturation compared to uninfected controls [25]. To determine whether ZIKV can infect and replicate in cells relevant to cardiac pathophysiology, we utilized fetal cardiac mesenchymal stromal cells (fcMSCs), which were previously isolated from fetal hearts of six to nine-weekold embryos [26]. Cardiac MSCs represent a major population of non-myocyte heart cells.…”

Section: Introductionmentioning

confidence: 99%

Characterization of zika virus infection of human fetal cardiac mesenchymal stromal cells

et al. 2020

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Zika virus (ZIKV) is a single-stranded RNA virus belonging to the family Flaviviridae. ZIKV predominantly enters cells using the TAM-family protein tyrosine kinase receptor AXL, which is expressed on a range of cell types, including neural progenitor cells, keratinocytes, dendritic cells, and osteoblasts. ZIKV infections have been associated with fetal brain damage, which prompted the World Health Organization to declare a public health emergency in 2016. ZIKV infection has also been linked to birth defects in other organs. Several studies have reported congenital heart defects (CHD) in ZIKV infected infants and cardiovascular complications in adults infected with ZIKV. To develop a better understanding of potential causes for these pathologies at a cellular level, we characterized ZIKV infection of human fetal cardiac mesenchymal stromal cells (fcMSCs), a cell type that is known to contribute to both embryological development as well as adult cardiac physiology. Total RNA, supernatants, and/or cells were collected at various time points post-infection to evaluate ZIKV replication, cell death, and antiviral responses. We found that ZIKV productively infected fcMSCs with peak (~70%) viral mRNA detected at 48 h. Use of an antibody blocking the AXL receptor decreased ZIKV infection (by~50%), indicating that the receptor is responsible to a large extent for viral entry into the cell. ZIKV also altered protein expression of several mesenchymal cell markers, which suggests that ZIKV could affect fcMSCs' differentiation process. Gene expression analysis of fcMSCs exposed to ZIKV at 6, 12, and 24 h post-infection revealed up-regulation of genes/pathways associated with interferonstimulated antiviral responses. Stimulation of TLR3 (using poly I:C) or TLR7 (using Imiquimod) prior to ZIKV infection suppressed viral replication in a dose-dependent manner. Overall, fcMSCs can be a target for ZIKV infection, potentially resulting in CHD during embryological development and/or cardiovascular issues in ZIKV infected adults.

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Section: Introductionmentioning

confidence: 99%

Characterization of zika virus infection of human fetal cardiac mesenchymal stromal cells

et al. 2020

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“…One limitation in our study is that we did not compare the survival of cartilage implants with and without Geltrex. In the future, it will be necessary to evaluate the effect of Adv-RFP-LOXL2 in implants without Geltrex to determine if its benefits are related to its ability to promote pluripotent stem cell differentiation and support chondrogenesis 21 , 22 .…”

Section: Discussionmentioning

confidence: 99%

“…These implants were surgically implanted subcutaneously in the backs of nude mice (3 implants/mouse). These explants were implanted with specialized matrix (Geltrex; ThermoFisher Scientific, Inc) as a basement membrane to promote a chondrogenic environment 21 , 22 . Geltre matrix is composed of laminin, collagen IV, entactin, and heparan sulfate proteoglycans.…”

Section: Methodsmentioning

confidence: 99%

“…Geltrex is lactose dehydrogenase-elevating virus (LDEV)-free and basement membrane matrix with very low growth factor activity. Geltrex helps to maintain pluripotent stem cells and does not adversely affect chondrogenic differentiation 21 . Furthermore, the combination of adenovirus with Geltrex was previously used to study endochondral ossification during porcine cartilage repair in vivo 22 .…”

Section: Methodsmentioning

confidence: 99%

“…We developed a model in which human TMJ cartilage was implanted in nude mice in combination with a specialized matrix of the basement membrane. This commercially available matrix, Geltrex, helps to maintain pluripotent stem cells without adversely affecting chondrogenic differentiation 21 . Previous studies also used the combination of adenovirus with Geltrex to study endochondral ossification during porcine cartilage repair in vivo 22 .…”

Section: Introductionmentioning

confidence: 99%

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LOXL2 promotes aggrecan and gender-specific anabolic differences to TMJ cartilage

Tashkandi

Alsaqer

Alhousami

et al. 2020

Sci Rep

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In the United States, 5–12% of adults have at least one symptom of temporomandibular joint (TMJ) disorders, including TMJ osteoarthritis (TMJ-OA). However, there is no chondroprotective agent that is approved for clinical application. We showed that LOXL2 is elevated in the regenerative response during fracture healing in mice and has a critical role in chondrogenic differentiation. Indeed, LOXL2 is an anabolic effector that attenuates pro-inflammatory signaling in OA cartilage of the TMJ and knee joint, induces chondroprotective and regenerative responses, and attenuates NF-kB signaling. The specific goal of the study was to evaluate if adenoviral delivery of LOXL2 is anabolic to human and mouse TMJ condylar cartilage in vivo and evaluate the protective and anabolic effect on cartilage-specific factors. We employed two different models to assess TMJ-OA. In one model, clinical TMJ-OA cartilage from 5 different samples in TMJ-OA cartilage plugs were implanted subcutaneously in nude mice. Adenovirus LOXL2 -treated implants showed higher mRNA levels of LOXL2, ACAN, and other anabolic genes compared to the adenovirus-Empty-treated implants. Further characterization by RNA-seq analysis showed LOXL2 promotes proteoglycan networks and extracellular matrix in human TMJ-OA cartilage implants in vivo. In order to evaluate if LOXL2-induced functional and sex-linked differences, both male and female four-month-old chondrodysplasia (Cho/+) mice, which develop progressive TMJ-OA due to a point mutation in the Col11a1 gene, were subjected to intraperitoneal injection with Adv-RFP-LOXL2 every 2 weeks for 12 weeks. The data showed that adenovirus delivery of LOXL2 upregulated LOXL2 and aggrecan (Acan), whereas MMP13 expression was slightly downregulated. The fold change expression of Acan and Runx2 induced by Adv-RFP-LOXL2 was higher in females compared to males. Interestingly, Adv-RFP-LOXL2 injection significantly increased Rankl expression in male but there was no change in females, whereas VegfB gene expression was increased in females, but not in males, as compared to those injected with Adv-RFP-Empty in respective groups. Our findings indicate that LOXL2 can induce specifically the expression of Acan and other anabolic genes in two preclinical models in vivo. Further, LOXL2 has beneficial functions in human TMJ-OA cartilage implants and promotes gender-specific anabolic responses in Cho/+ mice with progressive TMJ-OA, suggesting its merit for further study as an anabolic therapy for TMJ-OA.

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CRISPR-Knockout Screen Identifies Dmap1 as a Regulator of Chemically Induced Reprogramming and Differentiation of Cardiac Progenitors

Palano

Lim

et al. 2019

Stem Cells

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Direct in vivo reprogramming of cardiac fibroblasts into myocytes is an attractive therapeutic intervention in resolving myogenic deterioration. Current transgene‐dependent approaches can restore cardiac function, but dependence on retroviral delivery and persistent retention of transgenic sequences are significant therapeutic hurdles. Chemical reprogramming has been established as a legitimate method to generate functional cell types, including those of the cardiac lineage. Here, we have extended this approach to generate progenitor cells that can differentiate into endothelial cells and cardiomyocytes using a single inhibitor protocol. Depletion of terminally differentiated cells and enrichment for proliferative cells result in a second expandable progenitor population that can robustly give rise to myofibroblasts and smooth muscle. Deployment of a genome‐wide knockout screen with clustered regularly interspaced short palindromic repeats‐guide RNA library to identify novel mediators that regulate the reprogramming revealed the involvement of DNA methyltransferase 1‐associated protein 1 (Dmap1). Loss of Dmap1 reduced promoter methylation, increased the expression of Nkx2‐5, and enhanced the retention of self‐renewal, although further differentiation is inhibited because of the sustained expression of Cdh1. Our results hence establish Dmap1 as a modulator of cardiac reprogramming and myocytic induction. Stem Cells 2019;37:958–972

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Wnt/β-Catenin Stimulation and Laminins Support Cardiovascular Cell Progenitor Expansion from Human Fetal Cardiac Mesenchymal Stromal Cells

Cited by 21 publications

References 54 publications

Characterization of zika virus infection of human fetal cardiac mesenchymal stromal cells

Characterization of zika virus infection of human fetal cardiac mesenchymal stromal cells

LOXL2 promotes aggrecan and gender-specific anabolic differences to TMJ cartilage

CRISPR-Knockout Screen Identifies Dmap1 as a Regulator of Chemically Induced Reprogramming and Differentiation of Cardiac Progenitors

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