Supplementary MaterialsSupplementary information 41598_2018_28117_MOESM1_ESM. the skeletal muscle tissue can be considered as a central hinge of this disease. The muscle-pancreas axis was described for the first time by our group in 20111. In this study, we established a model in which primary beta-cells were 2′-Deoxycytidine hydrochloride treated with conditioned medium prepared from human primary myotubes obtained from vastus lateralis biopsies. We reported that human skeletal muscle cells produce and release myokines depending on their state of insulin sensitivity, with bimodal action depending on insulin resistance 2′-Deoxycytidine hydrochloride of the skeletal muscle cells used to condition culture medium1,2. Nevertheless, although all skeletal muscles share the same contractile function, they cannot be considered a homogenous organ from a metabolic point of view. The human body contains about 600 skeletal muscles, which can be classified in three main groups. Type I 2′-Deoxycytidine hydrochloride muscles (e.g. soleus) are mainly composed of type I fibers that are characterized by a slow ATP consumption rate and an oxidative metabolism able to generate enough ATP to cover energy needs during a long exercise3. Type II muscles (e.g. triceps brachii) are mainly composed of type II fibers and are highly fatigable. Type II fibers have a rate limiting step of glycolytic metabolism and therefore cannot generate enough ATP to cover the high ATP consuming rate of myosin heavy chain II during exercise of long duration3. The last group is composed of muscles made up of an approximately equivalent amount of type I and type II fibers (e.g. vastus lateralis)4. In the present work, we’ve established human types of skeletal muscle tissue cells isolated from type I and type II muscle groups and research their awareness to TNF-alpha induced insulin level of resistance. We have after that Rabbit polyclonal to smad7 investigated the way the muscle tissue type influences the profile of myokines secretion and their impact on beta-cells in order to identify new myokines implicated in fiber type specific muscle pancreas crosstalk. We show here for the first time, that skeletal muscle cells from biopsies with different fiber type composition present a unique gene expression and myokine signature. Moreover, the effect of human skeletal muscle cells on pancreatic beta-cells is usually fiber type specific, with both positive and negative effects depending on the level of insulin sensitivity. Finally we show that angiogenin (ANG) and osteoprotegerin (OPG) are triceps specific myokines that reduce apoptosis of beta-cells. These 2 myokines also prevent the apoptosis induced either by pro-inflammatory cytokines (cytomix: TNF-alpha, INFgamma and IL-1beta) or the unfavorable effect of insulin resistant conditioned medium from soleus skeletal muscle cells (TNF-S-CM). Morevover, OPG counteracts both the cytomix and TNF-S-CM negative effects on primary pancreatic beta-cells proliferation and insulin secretion. Results RNA sequencing (RNA-seq) approach reveals a unique signature in cells isolated from soleus and triceps biopsies In 2′-Deoxycytidine hydrochloride order to characterize the transcriptomes of biopsies and primary differentiated myotubes from soleus, triceps and vastus muscle, we established gene expression profiles using RNA-seq. The correlation of the overall gene expression within biopsies or myotubes is very high (spearman rho ~0.9) whereas it drops when comparing the biopsies with the myotubes (spearman rho ~0.5) (Supplementary Fig.?1). A principal component analysis (PCA) on RPKM values segregates well the biopsies from the differentiated myotubes (Fig.?1A, PC1). The soleus and the triceps biopsies form two distinct clusters while the vastus is usually more spread. This probably reflects the heterogeneous structure of this muscle type composed 2′-Deoxycytidine hydrochloride of both type I and II fibers (Supplementary Fig.?2, PC1 and PC2). The separation between the soleus and the triceps in induced myotubes is usually less evident probably because of the incomplete differentiation of the cultured cells (Supplementary Fig.?3). The comparison between induced myotubes MC-S and MC-T shows 2935 expressed genes that hit gene differentially.