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  • 1.
    Al-Khalili, Lubna
    et al.
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    de Castro Barbosa, T.
    Östling, J.
    Massart, J.
    Katayama, M.
    Nyström, A. -C
    Oscarsson, J.
    Zierath, J. R.
    Profiling of human myotubes reveals an intrinsic proteomic signature associated with type 2 diabetes2014Ingår i: Translational Proteomics, ISSN 2212-9634, Vol. 2, nr 1, s. 25-38Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    The development of insulin resistance and type 2 diabetes (T2D) involves a complex array of metabolic defects in skeletal muscle. An in vitro cell culture system excludes the acute effects of external systemic factors existing in vivo. Thus, we aimed to determine whether intrinsic differences in the protein profile exist in cultured myotubes derived from T2D versus normal glucose tolerant (NGT) healthy people. Applying two dimensional difference gel electrophoresis technology (2-D DIGE), the abundance of 47 proteins differed in myotubes derived from T2D patients versus NGT donors. Proteins involved in fatty acid and amino acid metabolism, TCA cycle, mitochondrial function, mRNA processing, DNA repair and cell survival showed higher abundance, while proteins associated with redox signaling (PARK7; Parkinson disease 7), glutathione metabolism (glutathione S-transferase, GST, isoforms T1, P1 and M2), and protein dynamics (heat shock protein, HSP, isoform B1 and 90A) showed reduced abundance in myotubes derived from T2D versus NGT donors. Consistent with our proteome analysis results, the level of total glutathione was reduced in myotubes obtained from T2D versus NGT donors. Taken together, our data provide evidence for intrinsic differences in the profile of proteins involved in energy metabolism, cellular oxidative stress, protein dynamics and gene regulation in myotubes derived from T2D patients. These differences thereby suggest a genetic or epigenetic influence on protein content level, which can be further investigated to understand the molecular underpinnings of T2D progression and lead to new therapeutic approaches.

  • 2.
    Al-Khalili, Lubna
    et al.
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Gillner, Karin
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Zhang, Ye
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Åstrand, Carolina
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Shokri, Atefeh
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Hughes-Brittain, Nanaaya
    McKean, Robert
    Robb, Brendan
    Chotteau, Véronique
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Characterization of Human CD133+Cells in Biocompatible Poly(l-lactic acid) Electrospun Nano-Fiber Scaffolds2016Ingår i: Journal of Biomaterials and Tissue Engineering, ISSN 2157-9083, E-ISSN 2157-9091, Vol. 6, nr 12, s. 959-966Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    CD133+ cells are potential myogenic progenitors for skeletal muscle regeneration to treat muscular dystrophies. The proliferation of human CD133+ stem cells was studied for 14 days in 3D biomimetic electrospun poly-L-lactic acid (PLLA) nano-fiber scaffolds. Additionally, the myogenic differentiation of the cells was studied during the last 7 days of the culture period. The cells were homogeneously distributed in the 3D scaffolds while colony formation and myotube formation occurred in 2D. After a lag phase due to lower initial cell attachment and an adaptation period, the cell growth rate in 3D was comparable to 2D after 7 and 14 days of culture. The expression of the stem cell (SC) marker PAX7 was 1.5-fold higher in 3D than 2D while the differentiation markers MyoG, Desmin and MyoD were only slightly changed (or remain unchanged) in 3D but strongly increased in 2D (12.6, 3.9, and 7.9-fold), and the myotube formation observed in 2D was absent in 3D. The marker expression during proliferation and differentiation, together with the absence of myotubes in 3D, indicates a better maintenance of stemness in 3D PLLA and stronger tendency for spontaneous differentiation in 2D culture. This makes 3D PLLA a promising biomaterial for the expansion of functional CD133+ cells.

  • 3. Garrido, Pablo
    et al.
    Salehzadeh, Firoozeh
    Duque-Guimaraes, Daniella E.
    Al-Khalili, Lubna
    Karolinska Institutet, Stockholm, Sweden.
    Negative regulation of glucose metabolism in human myotubes by supraphysiological doses of 17 beta-estradiol or testosterone2014Ingår i: Metabolism: Clinical and Experimental, ISSN 0026-0495, E-ISSN 1532-8600, Vol. 63, nr 9, s. 1178-1187Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Objective. Exposure of skeletal muscle to high levels of testosterone or estrogen induces insulin resistance, but evidence regarding the direct role of either sex hormone on metabolism is limited. Therefore, the aim of this study was to investigate the direct effect of acute sex hormone exposure on glucose metabolism in skeletal muscle. Materials/Methods. Differentiated human skeletal myotubes were exposed to either 17 beta-estradiol or testosterone and metabolic characteristics were assessed. Glucose incorporation into glycogen, glucose oxidation, palmitate oxidation, and phosphorylation of key signaling proteins were determined. Results. Treatment of myotubes with either 17 beta-estradiol or testosterone decreased glucose incorporation into glycogen. Exposure of myotubes to 17 beta-estradiol reduced glucose oxidation under basal and insulin-stimulated conditions. However, testosterone treatment enhanced basal palmitate oxidation and prevented insulin action on glucose and palmitate oxidation. Acute stimulation of myotubes with testosterone reduced phosphorylation of S6K1 and p38 MAPK. Exposure of myotubes to either 17 beta-estradiol or testosterone augmented phosphorylation GSK3 beta(ser9) and PKC delta(Thr505), two negative regulators of glycogen synthesis. Treatment of myotubes with a PKC specific inhibitor (GFX) restored the effect of either sex hormone on glycogen synthesis. PKC delta silencing restored glucose incorporation into glycogen to baseline in response to 17 beta-estradiol, but not testosterone treatment. Conclusion. An acute exposure to supraphysiological doses of either 17 beta-estradiol or testosterone regulates glucose metabolism, possibly via PKC signaling pathways. Furthermore, testosterone treatment elicits additional alterations in serine/threonine kinase signaling, including the ribosomal protein S6K1 and p38 MAPK.

  • 4. Pandya, J.
    et al.
    Venalis, P.
    Al-Khalili, Lubna
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Hossain, M.
    Stache, V.
    Lundberg, I. E.
    Malmström, V.
    Fasth, A.
    CD28null T cells are myotoxic to autologous muscle cells from patients with polymyositis2014Ingår i: Scandinavian Journal of Rheumatology, ISSN 0300-9742, E-ISSN 1502-7732, Vol. 43, s. 9-10Artikel i tidskrift (Övrigt vetenskapligt)
  • 5. Pandya, Jayesh M.
    et al.
    Venalis, Paulius
    Al-Khalili, Lubna
    KTH.
    Hossain, Mohammad Shahadat
    Stache, Vanessa
    Lundberg, Ingrid E.
    Malmström, Vivianne
    Fasth, Andreas E. R.
    CD4+and CD8+CD28(null) T Cells Are Cytotoxic to Autologous Muscle Cells in Patients With Polymyositis2016Ingår i: Arthritis & Rheumatology, ISSN 2326-5191, E-ISSN 2326-5205, Vol. 68, nr 8, s. 2016-2026Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Objective. Inflammatory T cell infiltrates in the skeletal muscle tissue of patients with polymyositis are dominated by CD28-negative effector (CD28(null)) T cells of both the CD4 and CD8 lineage. These cells are potentially cytotoxic, and the aim of the present study was to develop a fully autologous cell culture system in which to investigate the functional contribution of such CD28(null) T cells to myotoxicity. Methods. In vitro cocultures of autologous skeletal muscle cells and T cell subsets obtained from 5 polymyositis patients were performed. Myotoxicity of T cells was quantified by calcein release and flow cytometric analyses. T cell degranulation was blocked with concanamycin A. Specific blocking of perforin, cytokines, and HLA was performed using antibodies. Results. Both CD4+CD28(null) and CD8+CD28(null) T cells induced more muscle cell death than did their CD28+ counterparts. Differentiated muscle cells (myotubes) were more sensitive to T cell-mediated cell death than were their precursors (myoblasts). Both CD8+ and CD4+ CD28(null) T cells displayed perforin polarization toward muscle cells and secreted higher levels of granzyme B and interferon-gamma (IFN gamma) in coculture than did CD28+ T cells. The myotoxic effects of CD28(null) T cells were reduced upon the blocking of perforin, cytokines, and HLA. Addition of IFN gamma or tumor necrosis factor did not induce skeletal muscle cell death in the absence of T cells; however, it did up-regulate HLA expression on muscle cells. Conclusion. Myotoxicity of CD4+ and CD8+ CD28(null) T cells is mediated by directed perforin-dependent killing and can be further influenced by IFN gamma-induced HLA expression on muscle cells. The data suggest that CD28(null) T cells are key effector cells that contribute to the muscle cell damage in polymyositis.

  • 6. Pandya, Jayesh
    et al.
    Venalis, Paulius
    Al-Khalili, Lubna
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Hossain, Mohammad Shahadat
    Lundberg, Ingrid E.
    Malmström, Vivianne
    Fasth, Andreas E. R.
    CD28null T Cells from Polymyositis Patients Are Cytotoxic to Autologous Muscle Cells in Vitro Via Perforin-Dependent Mechanisms2014Ingår i: Arthritis & Rheumatology (Hoboken), ISSN 2326-5191, Vol. 66, s. S1193-S1193Artikel i tidskrift (Övrigt vetenskapligt)
  • 7. Venalis, P.
    et al.
    Pandya, J.
    Al-Khalili, Lubna
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Hossein, M. S.
    Stache, V.
    Lundberg, I. E.
    Malmström, V.
    Fasth, A. E. R.
    CD28NULL T CELLS KILL AUTOLOGOUS MUSCLE CELLS FROM POLYMYOSITIS PATIENTS IN VITRO BY PERFORIN-DEPENDENT MECHANISMS2014Ingår i: Annals of the Rheumatic Diseases, ISSN 0003-4967, E-ISSN 1468-2060, Vol. 73, s. 576-576Artikel i tidskrift (Övrigt vetenskapligt)
1 - 7 av 7
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