transforming-growth-factor-beta and Progeria

Topics: Diagnosis, Differential; DNA; Forkhead Box Protein M1; Forkhead Transcription Factors; Humans; Progeria; Prognosis; Transcription Factors; Transforming Growth Factor beta

Collagen type III (COL3) is one of the 3 major collagens in the body, and loss of expression or mutations in the

Topics: Aged; Cells, Cultured; Cellular Senescence; Collagen Type III; Extracellular Matrix; Humans; Infant; Mesenchymal Stem Cells; Myofibroblasts; Nanog Homeobox Protein; Progeria; Smad2 Protein; Smad3 Protein; Transforming Growth Factor beta

Cellular senescence as a result of organismal aging or progeroid diseases leads to stem cell pool exhaustion hindering tissue regeneration and contributing to the progression of age related disorders. Here we discovered that ectopic expression of the pluripotent factor NANOG in senescent or progeroid myogenic progenitors reversed cellular aging and restored completely the ability to generate contractile force. To elicit its effects, NANOG enabled reactivation of the ROCK and Transforming Growth Factor (TGF)-β pathways-both of which were impaired in senescent cells-leading to ACTIN polymerization, MRTF-A translocation into the nucleus and serum response factor (SRF)-dependent myogenic gene expression. Collectively our data reveal that cellular senescence can be reversed and provide a novel strategy to regain the lost function of aged stem cells without reprogramming to the pluripotent state. Stem Cells 2017;35:207-221.

Topics: Actins; Aged; Cell Differentiation; Cellular Senescence; Gene Expression Regulation; Genome, Human; Humans; Mesenchymal Stem Cells; Models, Biological; Muscle Development; Myofibroblasts; Nanog Homeobox Protein; Phenotype; Progeria; rho-Associated Kinases; Serum Response Factor; Signal Transduction; Trans-Activators; Transcription, Genetic; Transforming Growth Factor beta

Topics: Adolescent; Child, Preschool; Collagen; Elastin; Humans; Hyaluronic Acid; Male; Progeria; Transforming Growth Factor beta; Transforming Growth Factor beta1

Elastin and type IV collagen production are markedly elevated in fibroblasts derived from the skin of patients with Hutchinson-Gilford progeria (HGP). Fibroblasts from three affected children and their parents were compared to normal human skin fibroblasts with respect to elastin production as a function of different concentrations of calf serum and the cytokines, transforming growth factor-beta and basic fibroblast growth factor (TGF-beta 1, bFGF). In cultured fibroblasts from the parents of probands that were very high elastin producers (> 10(5) molecular equivalents/cell per h), at least one parent (mother) presented the same phenotype. Overproduction of elastin in culture could have been due to increased sensitivity of HGP strains to stimuli present in serum; however, relative stimulation of elastin production by calf serum in cell strains from HGP elastin over-producers was less than half the control strain. In most of the cultures examined, the responsiveness of elastin production to TGF-beta 1 was almost absent when compared to the response of normal fibroblasts. HGP strains with high elastin production modified conditioned medium to enhance elastin production in normal cells. These results suggest the presence, in HGP skin fibroblasts, of inheritance of high elastin production that is associated with accelerated aging.

Topics: Adolescent; Adult; Cells, Cultured; Child; Elastin; Female; Fibroblast Growth Factor 2; Fibroblasts; Humans; Infant; Male; Phenotype; Progeria; Skin; Skin Physiological Phenomena; Transforming Growth Factor beta

Fibroblasts from young patients exhibiting clinical features of progeroidal syndromes showed decreased biosynthesis of the small proteoglycan decorin. Cells in culture were metabolically labeled, and proteoglycans secreted into the medium were analyzed electrophoretically after immunoprecipitation with antibodies raised against decorin and biglycan. Fluorograms showed regularly a reduction to 15-30% of the normal amount of mature decorin and its core protein after chondroitin ABC lyase treatment. The size of the glycosaminoglycan chains was increased, but there was no obvious anomaly in the secretion kinetics of the mature proteoglycan. In addition, the patients' fibroblasts synthesized an increased amount of biglycan compared to control cells from healthy donors. Northern blot analysis clearly demonstrated a reduction by 85-94% in decorin mRNA, but biglycan mRNA was concomitantly increased, indicating that these alterations occur at the transcriptional level of protein expression. Transcription of decorin in fibroblasts from one of the patients was stimulated up to 3-fold by treatment with interleukin-1 beta. No response to interleukin-1 beta and transforming growth factor-beta was observed in the cells from another patient. In situ hybridization of cultured cells with an antisense decorin probe showed that decorin levels were reduced throughout the cell population. Surprisingly, subsequent examination of cells from one of the patients, now in mid-teenage, revealed a return to normal levels of decorin expression compared to age-matched controls. These studies suggest that, as in Marfan's syndrome where the primary defect concerns the fibrillin gene, reduced decorin expression contributes to the formation of an abnormal matrix and the pathogenesis of these disorders. They also indicate that this abnormality is likely to represent a secondary phenomenon which leads to a fault in the regulation of decorin gene transcription.

Topics: Base Sequence; Biglycan; Blotting, Northern; Cells, Cultured; Child, Preschool; Decorin; DNA; Electrophoresis, Polyacrylamide Gel; Extracellular Matrix Proteins; Female; Fibroblasts; Gene Library; Humans; In Situ Hybridization; Kinetics; Male; Molecular Sequence Data; Oligonucleotide Probes; Progeria; Proteoglycans; RNA, Messenger; Skin; Transforming Growth Factor beta

The basal levels of mRNAs encoding two metalloproteinases, collagenase and stromelysin, were increased as a function of in vitro serial subcultivation (cellular aging) of human fibroblasts. Procollagenase and prostromelysin synthesis and secretion were also greater in the old cultures (late passage). In contrast, the steady-state expression of mRNA for an inhibitor of metalloproteinases, tissue inhibitor of metalloproteinase-1 (TIMP-1), in late-passage cultures was lower than that in young cell cultures (early passage). Each mRNA was analyzed using total RNA preparations isolated from normal fibroblast cultures at different phases of the in vitro life span and from cultures derived from donors with the premature senescence syndromes characterized as Werner syndrome, progeria (Hutchinson-Gilford) syndrome, or Cockayne syndrome. In normal cell cultures expression of metalloproteinase mRNAs was increased after the culture had completed greater than 90% of the in vitro life span, and the reduction in TIMP-1 mRNA expression occurred after the culture had completed greater than 74% of the in vitro lifespan. In Werner syndrome cultures expression of metalloproteinase and TIMP-1 mRNAs was similar to the level of expression observed in late-passage cell cultures. Levels of metalloproteinase and TIMP-1 mRNA expression in progeria and Cockayne syndromes were similar to those of early-passage cell cultures. To determine if young and old cells were each responsive to mediators of metalloproteinase synthesis, cultures were treated with phorbol ester or cytokines. 12-O-tetradecanoylphorbol-13-acetate treatment increased the steady-state levels of all three mRNAs in young, old, and Werner syndrome cultures and increased procollagenase levels in all cultures. Early- and late-passage cell cultures also responded to cytokines. Interleukin-1 alpha treatment increased collagenase and stromelysin mRNA levels while transforming growth factor-beta reduced the steady-state levels of both transcripts. Neither cytokine affected the steady-state level of TIMP-1 mRNA. The results indicate that in vitro cellular aging is associated with changes in expression of mRNAs encoding proteins that mediate inflammatory responses and connective tissue remodeling.

Topics: Aging; Cells, Cultured; Cockayne Syndrome; Fibroblasts; Gene Expression Regulation; Glycoproteins; Humans; Interleukin-1; Matrix Metalloproteinase 3; Metalloendopeptidases; Microbial Collagenase; Phorbol Esters; Progeria; RNA, Messenger; Tissue Inhibitor of Metalloproteinases; Transforming Growth Factor beta; Werner Syndrome