Supplementary Figure 1. Increased expression of cell cycle pathway genes in insulin + Glut2 low cells of STZ-induced diabetic islets. A) random blood glucose measuers of STZ and vehicle treated MIP-GFP transgenic mice on day 5 after treatment. B) FPKM values for leading edge genes of cell cycle pathway increased in diabetic insulin + Glut2 low cells.
Supplementary Figure 2. Expansion of human diabetic PMP population and their increased proliferation capacity is maintained throughout multiple passaging. Frequencies and diameters of clonal spheres produced from individual healthy (n = 1) and diabetic (n = 3) donors in secondary (2 ) and tertiary (3 ) passages. The primary (1 ) spheres produced after 2 week culture of primary islet cells were handpicked, pooled, dissociated and cultured at clonal density (5000 cells/ml) to form secondary spheres (A). The same procedure was repeated on 2 spheres after two weeks to produce 3 spheres (B). The individual data from diabetic donors were combined and compared with the single non-diabetic control. Two way ANOVA showed a significant interaction of diabetes and sphere diameter in both passages (F 2 (2,231) = 20.35, p<0.0001, and F 3 (2,108) = 28.21, p<0.001) with a main effect of diabetes on the total number of spheres(f 2 (1,231) = 44.63, p<0.0001, and F 3 (1,108) = 22.07, p<0.001). N = 25-150 observed wells/group and values indicate means +/- SEMs.
Supplementary Figure 3. The proportion of hormone + cells produced from human PMP single colonies increases in diabetic condition. Single clonal spheres produced from three type 1 diabetic (D) and two control (C) human pancreatic islets were differentiated in serum containing media for 2 weeks and were stained for insulin (A) or glucagon (B) expression. (A) Clonal spheres from diabetic samples produced greater numbers of insulin + cells (One way ANOVA, F (4,53) = 5.024, p = 0017). Tukey post test revealed a significant increase of insulin + cell production by D2 and D3 spheres (p<0.05). (B) Spherical colonies from D1 and D3 produced significantly higher proportion of glucagon + cells (One way ANOVA, F (4,68) = 9.701, p<0.0001, Tukey post-test, p<0.05). Number of analyzed spherical colonies is 10-20 per sample in both experiments. Values indicate means+/-sems.
Supplementary Figure 4. PMP population expands in aged human islets. Dissociated pancreatic islet cells from 4 different donors (43, 45, 65, and 68 years old) were cultured at clonal density to form clonal spheres. The frequencies and diameters of spherical colonies were assessed for each sample. Values indicate means+/-sems
Supplementary Table 1. Log2 FDR values for comparing the KEGG and REACTOME defined biological pathways of insulin + Glut2 low versus insulin + Glut2 high cells from non-diabetic MIP-GFP transgenic islets at p<0.05 Pathway Glut2 low vs Glut2 high (CTRL) REACTOME Neuronal System 5.69048 KEGG Oxidative Phosphorylation 5.26629 REACTOME Respiratory Electron Transport ATP Synthesis by Chemiosmotic 5.19768 Coupling and Heat Production by Uncoupling Proteins REACTOME Metabolism Of Proteins 5.11695 REACTOME Transmission Across Chemical Synapses 5.05002 KEGG Parkinsons Disease 4.9294 REACTOME Respiratory Electron Transport 4.92474 REACTOME Regulation Of Gene Expression In Beta Cells 4.89525 KEGG Maturity Onset Diabetes Of The Young 4.87029 KEGG Huntingtons Disease 4.55676 KEGG Protein Export 4.39027 REACTOME Regulation Of Insulin Secretion 4.3749 REACTOME Tca Cycle and Respiratory Electron Transport 4.31096 REACTOME Synthesis Secretion and Inactivation Of GIP 4.28179 REACTOME Diabetes Pathways 4.27967 REACTOME Asparagine N Linked Glycosylation 4.20826 REACTOME Regulation Of Beta Cell Development 4.02694 REACTOME Integration Of Energy Metabolism 4.00822 KEGG_Alzheimers Disease 3.84465 REACTOME Unfolded Protein Response 3.83372 REACTOME Neurotransmitter Release Cycle 3.76846 REACTOME Glucose Metabolism 3.72381 REACTOME Gaba Synthesis Release Reuptake and Degradation 3.71421 REACTOME N Glycan Trimming In The ER and Calnexin Calreticulin Cycle 3.69859
REACTOME Incretin Synthesis Secretion and Inactivation 3.66705 REACTOME Post Translational Protein Modification 3.66355 REACTOME Activation Of Chaperone Genes by Xbp1s 3.6217 REACTOME Calnexin Calreticulin Cycle 3.56536 REACTOME Membrane Trafficking 3.40633 REACTOME Synthesis Secretion and Inactivation Of GLP1 3.38973 REACTOME SRP Dependent Co-translational Protein Targeting To Membrane 3.35397 KEGG Cardiac Muscle Contraction 3.24123 KEGG Valine Leucine and Isoleucine Degradation 3.18079 REACTOME Neurotransmitter Receptor Binding and Downstream Transmission 3.14851 In The Postsynaptic Cell REACTOME Cytosolic Sulfonation Of Small Molecules 3.14136 REACTOME Translation 3.11422 REACTOME Glutamate Neurotransmitter Release Cycle 3.09208 REACTOME Il-35 and GM-CSF Signaling -3.0395 KEGG Axon Guidance -3.0463 REACTOME Yap1 and Wwtr1 Taz Stimulated Gene Expression -3.0487 REACTOME PPARA Activates Gene Expression -3.0657 REACTOME Hyaluronan Metabolism -3.066 REACTOME Cell Extracellular Matrix Interactions -3.1112 REACTOME Class A Rhodopsin Like Receptors -3.126 REACTOME Peptide Ligand Binding Receptors -3.1398 REACTOME Antigen Activates B-Cell Receptor Leading To Generation Of -3.2276 Second Messengers REACTOME Initial Triggering Of Complement -3.2738 REACTOME Immunoregulatory Interactions Between A Lymphoid and A Non -3.2857 Lymphoid Cell KEGG Nod Like Receptor Signaling Pathway -3.3474
REACTOME GPVI Mediated Activation Cascade -3.3513 REACTOME Toll Receptor Cascades -3.3849 REACTOME Transcriptional Regulation Of White Adipocyte Differentiation -3.3979 REACTOME Rig I Mda5 Mediated Induction Of IFN- Alpha Beta Pathways -3.3987 REACTOME Glycosaminoglycan Metabolism -3.4209 REACTOME TRAF6 Mediated IRF7 Activation -3.4298 KEGG T Cell Receptor Signaling Pathway -3.4362 KEGG Natural Killer Cell Mediated Cytotoxicity -3.4549 REACTOME Signaling by Rho GTPases -3.4656 KEGG Rig I Like Receptor Signaling Pathway -3.4765 KEGG Leukocyte Transendothelial Migration -3.5146 REACTOME Chemokine Receptors Bind Chemokines -3.7265 REACTOME Platelet Activation Signaling and Aggregation -3.7561 KEGG TGF Beta Signaling Pathway -3.7629 KEGG Cytosolic DNA Sensing Pathway -3.7704 REACTOME Semaphorin Interactions -3.7869 KEGG Autoimmune Thyroid Disease -3.984 KEGG Primary Immunodeficiency -3.9875 KEGG JAK-STAT Signaling Pathway -3.995 KEGG Fc Epsilon RI Signaling Pathway -3.9961 KEGG Chemokine Signaling Pathway -4.1062 REACTOME Developmental Biology -4.1319 REACTOME Axon Guidance -4.1524 REACTOME Cell Surface Interactions At The Vascular Wall -4.1607 KEGG Asthma -4.1806 REACTOME Integrin Cell Surface Interactions -4.2036 REACTOME Signaling by ILS -4.2332 KEGG Fc Gamma R Mediated Phagocytosis -4.2894
KEGG Allograft Rejection -4.3225 REACTOME Interferon Alpha Beta Signaling -4.3533 KEGG Regulation Of Actin Cytoskeleton -4.5734 REACTOME Collagen Formation -4.6526 KEGG Graft Versus Host Disease -4.6828 REACTOME Interferon Signaling -4.74 KEGG B-Cell Receptor Signaling Pathway -4.7514 REACTOME Interferon Gamma Signaling -4.8852 KEGG Pathways In Cancer -4.8977 KEGG ECM Receptor Interaction -4.899 KEGG Complement and Coagulation Cascades -4.9069 REACTOME Immune System -5.0132 REACTOME Extracellular Matrix Organization -5.2199 KEGG Focal Adhesion -5.255 KEGG Toll Like Receptor Signaling Pathway -5.3245 REACTOME Innate Immune System -5.3355 REACTOME Hemostasis -5.4677 KEGG Intestinal Immune Network For IgA Production -5.5703 KEGG Leishmania Infection -5.8698 REACTOME Cytokine Signaling In Immune System -6.0399 KEGG Hematopoietic Cell Lineage -6.4271 KEGG Cytokine Cytokine Receptor Interaction -9.0316
Supplementary Table 2. KEGG and REACTOME defined pathways with significant alteration in insulin + glut2 low cells of STZ-induced diabetic islets. Pathway Glut2 low vs. Glut2 high Glut2 low vs. Glut2 high (Ctrl) (STZ) REACTOME Potassium Channels NA 3.29933 REACTOME Interaction Between L1 and Ankyrins NA 3.24749 REACTOME Amine Derived Hormones NA 3.22801 REACTOME Amine Ligand Binding Receptors NA 3.06795 REACTOME Cell Cycle NA 3.05909 KEGG P53 Signaling Pathway NA 3.05151 REACTOME Inflammasomes NA -3.49665 REACTOME NCAM1 Interactions NA -4.1326 Values are log2 FDR at p<0.05