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WO2018125019A2 - Utilisation de certains miarn destinés au diagnostic et au traitement de maladies associées à l'insuline - Google Patents

Utilisation de certains miarn destinés au diagnostic et au traitement de maladies associées à l'insuline Download PDF

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WO2018125019A2
WO2018125019A2 PCT/TR2017/050737 TR2017050737W WO2018125019A2 WO 2018125019 A2 WO2018125019 A2 WO 2018125019A2 TR 2017050737 W TR2017050737 W TR 2017050737W WO 2018125019 A2 WO2018125019 A2 WO 2018125019A2
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insulin
cells
disease
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WO2018125019A3 (fr
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Selda OKTAYOĞLU
Ediz COŞKUN
Merve ERCIN
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Istanbul Ueniversitesi
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    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
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    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
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    • C12N5/0676—Pancreatic cells
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    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
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    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/178—Oligonucleotides characterized by their use miRNA, siRNA or ncRNA

Definitions

  • the present invention relates to use of certain microRNAs in the diagnosis and treatment of insulin-related diseases such as diabetes and obesity.
  • the invention further relates to use of said microRNAs in the cellular treatment of the insulin-related diseases such as diabetes and obesity, by allowing pancreatic ⁇ -cells to be formed via delivery of microRNAs to stem cells in vitro, and transplanting these cells to the patients.
  • pancreas is a mixed organ consisting of exocrine and endocrine parts.
  • the exocrine part secretes digestive enzymes
  • the endocrine part known as Langerhans islets secretes different hormones.
  • Each islet consists of a large number of specialized endocrine cells and these cells separate from each other due to their specific hormones.
  • An insulin-secreting ⁇ -cell, glucagon- secreting a-cell, somatostatin-secreting ⁇ -cell, and pancreatic polypeptide-secreting PP cells are the four major types of specialized cell types that constitute the islet.
  • the present invention includes at least one RNA selected from the group consisting of miR-18a, miR-19b, miR-30d, miR-124, miR-146a, miR-184, miR-335, miR-433-5p and miR-503, or a DNA, mRNA or IncRNA molecule thereof, for use in treatment of the insulin-related diseases.
  • Said insulin-related disease may be selected from the group consisting of prediabetes, Type I diabetes mellitus, Type II diabetes mellitus, metabolic syndrome, obesity, lactose intolerance, fructose intolerance, galactosemia, glycogen storage disease, insulin resistance syndrome, syndrome X, retinopathy, neuropathy, nephropathy, foot ulcers, hypertension, hyperlipidemia, metabolic syndrome, gall bladder diseases, osteoarthritis, cardiovascular diseases, stroke, sleep apnea, liver disease, asthma, respiratory distress, menstruation disorders, musculoskeletal system disorders, skin diseases, polycystic ovarian syndrome, immune system disorders, Alzheimer's disease and the other types of dementia, Parkinson's disease and other Parkinson's- related diseases, Huntington's disease, amyotrophic lateral sclerosis (ALS], Batten disease, motor neuron disease, spinal muscular atrophy, Prion disease, spinocerebellar ataxia, lewy body dementia, multiple sclerosis (MS], neuropathy and Friedrich ataxi
  • Molecules of the invention may be in combination with at least one additional active substance selected from antidiabetic, anti-obesity and anti-inflammatory agents.
  • Said antidiabetic agent may be selected from the group consisting of insulin analogs, insulin sensitizers, insulin secretagogues, aldose reductase inhibitors, alpha glucosidase inhibitors, amylin analogs, peptide analogs, sodium glucose transporter 2 (SGLT] inhibitors and glucosuric agents.
  • Said anti-obesity agent may be selected from the group consisting of 4-methyl amphetamine, amfecloral, amfepentorex, amfepramon, aminorex, amphetamine, atomoxetine, benfluorex, benzphetamine, bupropion, cathine, cathinone, chlorphentermine, cyclazindol, clobenzorex, cloforex, clominorex, clotermine, dexfenfluramine, dextroamphetamine, dexmethylphenidate, difemetorex, dimethylcathinone, difemethoxydine, ephedrine, ephedra, ethylamphetamine, etolorex, fenbutrazate, fencamfamin, fenethylline, fenfluramine, fenproporex, fludorex, fluminorex, furfenorex, indan
  • Said anti-inflammatory agent may be selected from the group consisting of pyrazolone/pyrazolidines, salicylates, acetic acid derivatives, oxicams, propionic acid derivatives, N-arylanthranilic acids and coxibs.
  • Molecule of the invention may be presented in a pharmaceutical formulation.
  • the formulation may be in a dosage form that may be administered to a patient orally, rectally, vaginally, intratumorally, subcutaneously, intracutaneously, intravenously, intracerebroventricularly, intramuscularly, intra-arterially, intrathecally, intranasally, interperitoneally, parenterally, topically, or by means of medical devices.
  • the formulation is suitable to be targeted to, or to be carried by, the cells via nanoparticles, liposomes or other carriers.
  • the invention in another aspect, relates to a method for obtaining pancreatic ⁇ -cells in vitro, said method comprising treatment of mesenchymal stem cells with at least one microRNA selected from miR-18a, miR-19b, miR-30d, miR-124, miR-146a, miR-184, miR-335, miR-433-5p and miR- 503, or a DNA, mRNA or a related IncRNA thereof.
  • pancreatic mesenchymal stem cells may be treated with HDAC (Histone deacetylase] inhibitor and/or glucose.
  • Said HDAC inhibitor is preferably valproic acid or a pharmaceutically acceptable salt thereof.
  • HDAC inhibitor has a concentration ranging between 0.5 mM to 5 mM.
  • glucose may have a concentration ranging between 5 mM to 50 mM.
  • the invention relates to pancreatic ⁇ -cells obtained from the method above for use in the treatment of the insulin-related diseases.
  • Said pancreatic ⁇ -cells may be presented in combination with at least one microRNA selected from the group consisting of miR-18a, miR-19b, miR-30d, miR-124, miR-146a, miR-184, miR-335, miR-433-5p and miR-503, or a DNA, mRNA or one or more related IncRNA.
  • Said ⁇ -cells may also be provided in combination with at least one additional agent selected from antidiabetic, anti-obesity and anti-inflammatory agents.
  • the invention includes the case in which said ⁇ -cells are provided in a pharmaceutical composition comprising at least one excipient
  • Said pharmaceutical composition is suitable to be targeted to, or to be carried by, the cells via the carriers selected from heparin, lactic acid based polymers, polyesters, hydrogels, biopolymer films, extravascular compartments, intravascular compartments, alginate, poly(hydroxyethylmethacrylate-methyl methacrylate], agarose, acrylonitrile copolimers, chitosan, and PEG nanoparticles and liposomes.
  • the invention in another aspect, relates to a method for in vitro diagnosis of the insulin-related diseases, said method comprising use of at least one microRNA selected from the group consisting of miR-18a, miR-19b, miR-30d, miR-124, miR-146a, miR-184, miR-335, miR-433-5p and miR-503, or a DNA, mRNA or one or more related IncRNA as a biomarker.
  • Diseases diagnosed herein are as listed above.
  • the diagnosis method comprises detecting at least one of said biomarkers in a sample selected from the group consisting of blood, plasma, serum, milk, bronchoalveolar fluid and cerebrospinal fluid.
  • the invention relates to a kit for in vitro diagnosis of the insulin-related diseases, preferably the above-mentioned diseases, the kit comprising a biomarker which comprises at least one microRNA selected from the group consisting of miR-18a, miR-19b, miR-30d, miR-124, miR-146a, miR-184, miR-335, miR-433-5p and miR-503, or a DNA, mRNA or IncRNA.
  • the invention provides a kit for in vitro amplification of pancreatic ⁇ -cells, including at least one microRNA selected from miR-18a, miR-19b, miR-30d, miR-124, miR-146a, miR-184, miR-335, miR-433-5p and miR-503, or a DNA, mRNA or IncRNA thereof.
  • a kit for in vitro amplification of pancreatic ⁇ -cells including at least one microRNA selected from miR-18a, miR-19b, miR-30d, miR-124, miR-146a, miR-184, miR-335, miR-433-5p and miR-503, or a DNA, mRNA or IncRNA thereof.
  • Figure 1 shows the effect of VP A of 0.75-3 mM on PI-MSC viability as % Control.
  • Figure 2 shows the effect of 25mM of Glucose, ImM of VP A, and 25 mM of Glucose+ 1 mM of VP A on the ⁇ -cell differentiation at the end of days 10, 20 and 30. Histogram plots showing flow cytometric insulin + cell number (%, Fluorescence intensity] (A], a graphic showing the analysis results (B], and mean ⁇ SEM values are provided. *p ⁇ 0,05, **p ⁇ 0,01, ***p ⁇ 0,001 vs. control.
  • Figure 3 shows the effect of experimental conditions applied to PI-MSCs for 20 days on mRNA production. A graphic representing the analysis results of mRNA levels (fold increase] (A] and mean ⁇ SEM values (B] are provided. *p ⁇ 0,05; **p ⁇ 0,01 and ***p ⁇ 0,001 vs. control.
  • Figure 4 shows microscopic photographs of the control group received complete medium (a] and the group received glucose (b] at the end of 20-day experiment period.
  • the upper panel is magnified 4X
  • the middle panel is magnified 10X
  • the lower panel is magnified 2 OX. *shows the stem cell colony.
  • Figure 5 shows microscopic photographs of the group received VPA (a] and the group received VPA+glucose (b] at the end of 20-day experiment period.
  • the upper panel is magnified 4X
  • the middle panel is magnified 10X
  • the lower panel is magnified 2 OX. *shows the stem cell colony and ⁇ shows a ⁇ -cell like round cell.
  • Figure 6 shows the effect of experimental conditions applied to PI-MSCs for 20 days on ROS production.
  • Dot plots showing flow cytometric DCF Fluorescence intensity (Fold Increase] (A], a graphic representing the analysis results (B] and mean ⁇ SEM values are provided. **p ⁇ 0,01 and ***p ⁇ 0,001 vs. control, ##p ⁇ 0,01 and ###p ⁇ 0,001 vs. group received glucose.
  • Figure 7 shows insulin amounts (ng/ ⁇ g protein] released from the cells as a result of 5.5 mM of glucose and 25 mM glucose administration. **p ⁇ 0,01 and ***p ⁇ 0,001 vs. control group.
  • Figure 8 demonstrates graphics showing OCT3/4, c-Myc and Nanog protein bands and their intensities in cytoplasmic (left panel] and nuclear (right panel] fractions of PI-MSCs. *p ⁇ 0,05, **p ⁇ 0,01, ***p ⁇ 0,001 vs. control, ##p ⁇ 0,01; ###p ⁇ 0,001 vs. group received glucose, and +p ⁇ 0,05 vs. group received valproic acid.
  • Figure 9 demonstrates graphics showing HDAC1 and HDAC3 protein bands and their intensities in cytoplasmic (left panel] and nuclear (rightpanel] fractions of PI-MSCs. **p ⁇ 0,01, ***p ⁇ 0,001 vs. control, ##p ⁇ 0,01; ###p ⁇ 0,001 vs. group received glucose.
  • Figure 10 demonstrates graphics showing H4-K12 Ace, H3-K9 Ace and H3-K9 TriMe protein bands of PI-MSCs, and band intensities thereof.
  • Figure 11 shows the changes in the expression levels of some miRNAs in the control group and the glucose-treated group. A statistically significant increase was found only in miRNAs that were above the blue line, while red circles show miRNAs, the gene expression levels of which are increased (p ⁇ 0,05].
  • Figure 12 shows the changes in the expression levels of some miRNAs in the control group and the VPA-treated group. Red circles show miRNAs, the gene expression levels of which are increased, whereas green circles show miRNAs, the gene expression levels of which are decreased. A statistically significant increase was found only in miRNAs that were above the blue line (p ⁇ 0,05).
  • Figure 13 shows the changes in the expression levels of some miRNAs in the control group and the VPA+glucose-treated group. Red circles show miRNAs, the gene expression levels of which are increased, whereas green circles show miRNAs, the gene expression levels of which are decreased. A statistically significant increase was found only in miRNAs that were above the blue line (p ⁇ 0,05).
  • Figure 14 shows the changes in the expression levels of some miRNAs in the glucose-treated group and the VPA+glucose-treated group. Red circles show miRNAs, the gene expression levels of which are increased, whereas green circles show miRNAs, the gene expression levels of which are decreased. A statistically significant increase or decrease was found only in miRNAs that were above the blue line (p ⁇ 0,05].
  • Figure 15 shows the changes in the expression levels of some miRNAs in the VPA+glucose-treated group vs. VPA-treated group. Red circles show miRNAs, the gene expression levels of which are increased, whereas green circles show miRNAs, the gene expression levels of which are decreased.
  • Figure 16 shows the grade for gene expression level of miRNAs in each group. Gene expression levels increase from green to red.
  • the invention relates to at least one RNA selected from the group consisting of miR- 18a, miR-19b, miR-30d, miR-124, miR-146a, miR-184, miR-335, miR-433-5p and miR-503, or a DNA, mRNA or IncRNA thereof, for use in the treatment of the insulin-related diseases, preferably diabetes and obesity.
  • the present invention relates to use of at least one RNA selected from the group consisting of miR-18a, miR-19b, miR-30d, miR-124, miR-146a, miR-184, miR-335, miR-433-5p and miR-503, or a DNA, mRNA or IncRNA thereof in the treatment of the insulin-related diseases, preferably diabetes and obesity as a medicament in combination with antidiabetic/anti-obesity drugs.
  • the invention relates to use of at least one RNA selected from the group consisting of miR-18a, miR-19b, miR-30d, miR-124, miR-146a, miR-184, miR-335, miR-433-5p and miR-503, or a DNA, mRNA or IncRNA thereof, alone or in combination with other antidiabetic, anti-obesity and anti-inflammatory drugs, in order to produce new ⁇ -cells in the cellular treatment of the insulin-related diseases, preferably diabetes and obesity. It is also suggested that more efficient cellular treatment methods can be developed by adding said molecules to the current ⁇ -cell differentiation protocols.
  • the present invention relates to use of at least one RNA selected from the group consisting of miR-18a, miR-19b, miR-30d, miR-124, miR-146a, miR-184, miR-335, miR-433-5p and miR-503, or a DNA, mRNA or IncRNA thereof, alone or in combination, or together with known biomarkers in the diagnosis of the insulin-related diseases, preferably diabetes and obesity.
  • mRNA used in the present invention refers to messenger RNAs. The mRNA molecules transfer the genetic information obtained from DNA to the ribosome.
  • the phrases "mRNA” and “messenger RNA” used in the present invention have the same meaning and are used interchangeably.
  • RNA used in the present invention refers to a small and non-coding RNA molecule. These molecules may be used for RNA silencing and regulation of post-transcriptional gene expression.
  • micro RNA and “miRNA” have the same meaning and may be used interchangeably.
  • IncRNA used in the present invention refers to long non-coding RNAs. These RNAs are transcripts that do not code for protein and have a length of more than 200 nucleotides.
  • lnc RNA and “long non-coding RNA” used in the present invention have the same meaning and may be used interchangeably.
  • diabetes mellitus used in the present invention refers to type I diabetes mellitus and/or type II diabetes mellitus.
  • insulin-related diseases used in the present invention refers to various neurodegenerative diseases such as lactose intolerance, fructose intolerance, galactosemia, glycogen storage disease, insulin resistance syndrome, syndrome X, retinopathy, nephropathy, foot ulcers, hypertension, hyperlipidemia, metabolic syndrome, gall bladder diseases, osteoarthritis, cardiovascular diseases, stroke, sleep apnea, liver disease, asthma, respiratory distress, menstruation disorders, musculoskeletal system disorders, skin diseases, polycystic ovarian syndrome, immune system disorders, diabetes mellitus, obesity and Alzheimer's disease and the other types of dementia, Parkinson's disease and other Parkinson's-related diseases, Huntington's disease, amyotrophic lateral sclerosis (ALS], Batten disease, motor neuron disease, spinal muscular atrophy, Prion disease, spinocer
  • ALS amyotrophic
  • MiR-184 is important for the regulation of ⁇ -cell mass increase i.e. compensation, in the case of insulin resistance.
  • Ago2 which is the target of miR-184 in pancreatic islets, is part of the RNA- induced silencing complex required for targeting mRNAs. Loss of Ago2 blocks the compensatory increase of ⁇ -cells, which develops in response to insulin resistance by increasing the expression of miR-375 targets.
  • miR-184 was suppressed in the islets of insulin resistant mice and humans, resulting in increased expression of Ago2, thereby compensatory increase of ⁇ -cells being suppressed.
  • Ago2 levels are decreased and compensatory increase of ⁇ -cells is suppressed.
  • MiR-335 is associated with ⁇ -cell function, rather than ⁇ -cell differentiation.
  • islets were isolated from Goto Kakizaki (GK] rats which were used as a type 2 diabetes mellitus, a ⁇ -cell dysfunction. It has been determined that miR-335 expression is increased in these islets and that their targets are mRNAs of Stxbpl, Sytll and Snap25 which are molecules associated with insulin exocytosis (Esguerra et al., 2011].
  • overexpression of miR-335 leads to a decrease in glucose-induced insulin secretion and a decrease in depolarization-induced insulin exocytosis.
  • VPA-treated groups in which we detected ⁇ -cell differentiation, unresponsiveness to glucose- induced insulin secretion is parallel to the increase in miR-335 expression.
  • MiR-30d is associated with pancreas development, whereas miR-19b is associated with HDCAs.
  • MiR-30 family are produced at high rates in the human fetal pancreas and are associated with epithelial-mesenchymal transition. These miRNAs inhibit mesenchymal mRNAs such as vimentin and Snaill and allow pancreatic mesenchymal stem cells to transform into insulin producing cells. It has been shown that miR-30d, a member of this family, regulates the ⁇ - cell transcription factor MafA, butdoes not affect Pdx-1 and NeuroDl. MiR-30d increased the MafA level and the transcription of the insulin gene.
  • MiR-19b has been reported to show an increase in acute myeloid leukemia cells treated with Vorinostat, a class I and II HDAC inhibitor. However, there was no study showing the relationship of miR-19b with HDAC except this study. It has also been shown that miR-19b is expressed at high levels in pancreatic progenitor cells and that this miRNA targets the 3 'UTR region of NeuroDl mRNA, thereby reducing the protein and mRNA levels of this transcription factor. It has also been shown that MiR- 19b inhibits insl expression in MIN6 cells, does not affect ins2, and has little effect on the proliferation of pancreatic progenitor cells.
  • MiR-124 is one of the best characterized and most abundantly expressed neuronal miRNAs. Overexpression of MiR-124 results in an increase in the expression of neuronal markers, and thus, neurite growth, indicative of neuronal differentiation, has also been shown to manifest itself as morphological changes. In some studies conducted in vertebrates, miR-124 has been identified as a stimulant for neuronal differentiation and as an inhibitor of self-renewal occurred in progenitor cells. However, it is not known how miR-124 directs neurogenesis in mesenchymal stem cells. In the study of the present invention, findings which suggest that miR-124 mediates differentiation of glucose-treated PI-MSCs into ⁇ -cells, were found for the first time in the literature.
  • the invention in another aspect, relates to a method for the formation of pancreatic ⁇ -cell suitable for the treatment of insulin-related diseases, preferably diabetes and obesity, comprising in vitro administration of at least one RNA selected from miR-18a, miR-19b, miR-30d, miR-124, miR-146a, miR-184, miR-335, miR-433-5p and miR-503, or a DNA, mRNA or IncRNA thereof, to the stem cells.
  • RNA selected from miR-18a, miR-19b, miR-30d, miR-124, miR-146a, miR-184, miR-335, miR-433-5p and miR-503, or a DNA, mRNA or IncRNA thereof, to the stem cells.
  • the invention relates to use of pancreatic ⁇ -cells obtained by co-administration of at least one RNA selected from the group consisting of miR-18a, miR-19b, miR-30d, miR-124, miR-146a, miR-184, miR-335, miR-433-5p and miR-503, or a DNA, mRNA or IncRNA thereof to the stem cells with at least one other antidiabetic, anti-obesity or anti-inflammatory agent, for the treatment of the insulin-related diseases, preferably for the treatment of diabetes and obesity.
  • at least one RNA selected from the group consisting of miR-18a, miR-19b, miR-30d, miR-124, miR-146a, miR-184, miR-335, miR-433-5p and miR-503, or a DNA, mRNA or IncRNA thereof to the stem cells with at least one other antidiabetic, anti-obesity or anti-inflammatory agent, for the treatment of the insulin-related diseases, preferably for the treatment
  • the invention relates to use of pancreatic ⁇ -cells obtained by in vitro administration of at least one RNA selected from the group consisting of miR-18a, miR-19b, miR- 30d, miR-124, miR-146a, miR-184, miR-335, miR-433-5p and miR-503, or a DNA, mRNA or IncRNA thereof, to the stem cells in combination with at least one other antidiabetic, anti-obesity or anti-inflammatory agent in the treatment of the insulin-related diseases, preferably in the treatment of diabetes and obesity.
  • RNA selected from the group consisting of miR-18a, miR-19b, miR- 30d, miR-124, miR-146a, miR-184, miR-335, miR-433-5p and miR-503, or a DNA, mRNA or IncRNA thereof, to the stem cells in combination with at least one other antidiabetic, anti-obesity or anti-inflammatory agent in the treatment of the insulin-related diseases,
  • the present invention also relates to a method for use in the diagnosis of the insulin-related diseases, preferably diabetes and obesity, comprising detecting the presence of at least one of biomarker comprising at least one RNA selected from the group consisting of miR-18a, miR-19b, miR-30d, miR-124, miR-146a, miR-184, miR-335, miR-433-5p and miR-503, or a DNA, mRNA or IncRNA thereof, in all body fluids such as blood, plasma, serum, milk, bronchoalveolar fluid and cerebrospinal fluid.
  • biomarker comprising at least one RNA selected from the group consisting of miR-18a, miR-19b, miR-30d, miR-124, miR-146a, miR-184, miR-335, miR-433-5p and miR-503, or a DNA, mRNA or IncRNA thereof, in all body fluids such as blood, plasma, serum, milk, bronchoalveolar fluid and cerebrospinal fluid
  • Said molecules may be determined by precise molecular methods such as Southern Blot, Northern Blot, PCR, RT-PCR, qRT-PCR, microarray and sequencing and the methods that may be used are not limited to said techniques.
  • Antidiabetic agent used in the present invention means agents used in the treatment of diabetes mellitus, whereas "anti-obesity agent” refers to agents used in the treatment of obesity. All molecules (alone or in combination], all the related RNAs and DNAs, which were described in the present invention and detailed above can be used in cellular therapy in conjunction with antidiabetic, anti-obesity or anti-inflammatory drugs.
  • Said antidiabetic agents may be selected from insulin analogs, insulin sensitizers, insulin secretagogues, aldose reductase inhibitors, alpha glucosidase inhibitors, amylin analogs, peptide analogs, sodium glucose transporter 2 (SGLT] inhibitors and glucosuric agents.
  • Insulin that can be used in combination with the molecules listed above in the context of the present invention may be selected from the group consisting of insulin, insulin lispro, insulin aspart, insulin glulisine, insulin zinc, isophane insulin, insulin glargine, insulin detemir.
  • Insulin sensitizers that can be used in combination with the molecules listed above in the context of the present invention may be selected from the group consisting of metformin, phenformin, buformin, siglitazone, darlitazone, englitazone, lobeglitazone, netoglitazone, rivoglitazone, aleglitazar, saroglitazar, tesaglitazar, rosiglitazone, pioglitazone, and troglitazone.
  • Insulin secretagogues that can be used in combination with the molecules listed above in the context of the present invention may be selected from the group consisting of acetohexamide, carbutamide, chlorpropamide, metahexamide, tolbutamide, tolazamide, glibenclamide, glibornuride, glicetanyl, gliclazide, gliflumide, glipizide, gliquidone, glisoxepide, glyclopyramide, glimepiride, repaglinide, mitiglinide, exenatide, liraglutide, taspoglutide, albiglutide, lixisenatide, dulagutide, semaglutide, alogliptin, anagliptin, gemigliptin, linagliptin, omarigliptin, saxagliptin, sitagliptin, tenegliptin, vildagliptin, fasiglif
  • Aldose reductase inhibitors that can be used in combination with the molecules listed above in the context of the present invention may be selected from the group consisting of epalrestat, fidarestat, ranirestat, tolrestat, and zenarestat.
  • Alpha glucosidase inhibitors that can be used in combination with the molecules listed above in the context of the present invention may be selected from the group consisting of miglitol, acarbose, and voglibose.
  • Amylin analog that can be used in combination with the molecules listed above in the context of the present invention is pramlintide.
  • Sodium glucose transporter 2 (SGLT] inhibitors that can be used in combination with the molecules listed above in the context of the present invention may be selected from the group consisting of canaglifozin, dapagliflozin, empagliflozin, remogliflozin, sergliflozin, and tofogliflozin.
  • the antidiabetic agent may also be benfluorex or bromocriptine.
  • Said anti-obesity agents may be selected from the group consisting of 4-methyl amphetamine, amfecloral, amfepentorex, amfepramon, aminorex, amphetamine, atomoxetine, benfluorex, benzphetamine, bupropion, cathine, cathinone, chlorphentermine, cyclazindol, clobenzorex, cloforex, clominorex, clotermine, dexfenfluramine, dextroamphetamine, dexmethylphenidate, difemetorex, dimethylcathinone, difemethoxydine, ephedrine, ephedra, ethylamphetamine, etolorex, fenbutrazate, fencamfamin, fenethylline, fenfluramine, fenproporex, fludorex, fluminorex, furfenorex, indan
  • Said anti-inflammatory agents may be selected from the general groups consisting of pyrazolone/pyrazolidines, salicylates, acetic acid derivatives, oxicams, propionic acid derivatives, N-arylanthranilic acids, coxibs and the other agents.
  • Said anti-inflammatory agents may be selected from the groups consisting of aminophenazone, ampyrone, clophenazone, famprofazone, feprazone, kebuzone, metamizole, mofebutazone, morazone, nifenazone, oxyphenbutazone, phenazone, phenylbutazone, propyphenazone, sulfinpyrazone, suxibuzone, acetylsalicylic acid, aloxypyrine, benorilate, carbasalate calcium, diflunisal, ethenzamide, guacetical, magnesium salicylate, methyl salicylate, salsalate, salicylamide, salicylic acid, sodium salicylate, aceclofenac, acemetacin, alclofenac, amfenac, bendazac, bromfenac, bumadizone, bufexamac, diclofenac, difenpiramide, etodolac
  • the invention relates to pharmaceutical compositions comprising ⁇ -cells obtained by the methods according to the invention.
  • Said pharmaceutical compositions may contain at least one other excipient in addition to the beta cells.
  • the pharmaceutical compositions according to the invention may contain at least one other active ingredient in addition to the ⁇ - cells.
  • the other active ingredient may be selected from the antidiabetic or anti-obesity agents described in detail above, or a combination thereof.
  • the other active agent may be formulated together with ⁇ -cells, or it may be formulated separately and administered to the patient simultaneously, sequentially or at different times.
  • compositions according to the present invention are administered to the culture medium in vitro to form beta cells, these cells can be injected intravenously.
  • Formulations suitable for injection may be formulated using a sterile solvent or any pharmaceutically acceptable carrier.
  • the pharmaceutically acceptable carrier may be selected from sterile water, saline, or the existing cell culture media within the scope of state of the art, but is not limited thereto.
  • compositions according to the present invention may be administered parenterally in the form of an injectable formulation.
  • Formulations suitable for injection may be formulated using a sterile solvent or any pharmaceutically acceptable carrier.
  • the pharmaceutically acceptable carrier may be selected from sterile water, saline, or the existing cell culture media within the scope of state of the art, but is not limited thereto.
  • the invention relates to a method of ⁇ -cell differentiation, comprising a step of obtaining ⁇ -cells by administrating a HDAC (Histone deacetylase] inhibitor and glucose to pancreatic mesenchymal stem cells.
  • HDAC Histone deacetylase
  • valproic acid or a pharmaceutically acceptable salt thereof, such as sodium valproate is used as the HDAC inhibitor.
  • the HDAC inhibitor is used in a range of 0.5 mM to 5 mM, preferably in a range of 0.75 mM to 3 mM, particularly preferably in a range of 1 mM to 2 mM.
  • the ⁇ -cell differentiation protocol uses glucose in a range of 5 mM to 50 mM, preferably in a range of 10 nM to 40 mM, particularly preferably in a range of 20 mM to 30 mM.
  • the invention relates to pharmaceutical formulations comprising at least one RNA selected from the group of miR-18a, miR-19b, miR-30d, miR-124, miR-146a, miR-184, miR- 335, miR-433-5p ve miR-503, or a DNA, mRNA or IncRNA thereof.
  • compositions according to the invention may, in addition to said RNAs, contain at least one pharmaceutically acceptable excipient and/or additional active ingredient
  • the other active agent mentioned herein may be an antidiabetic agent or an anti-obesity agent or anti-inflammatory agent Details of these agents are given in the context of the invention.
  • the formulations according to the invention may be prepared in the form of any of the existing dosage forms known in the art of composition. Said dosage form may be administered to a patient orally, rectally, vaginally, intratumorally, subcutaneously, intracutaneously, intravenously, intracerebroventricularly, intramuscularly, intraarterially, intratracheally, interperitoneally, parenterally, topically, or by means of medical devices.
  • the formulations according to the present invention can be suitably formulated to be targeted to, or to be carried by, the cells via nanoparticles, liposomes and the other similar carriers, or they can be administered by said routes.
  • the formulations according to the present invention may be formulated in a manner suitable for administration to the patient by nasal, spray, oral, aerosol, rectal or vaginal route of administration, or they can be administered to the patient by said routes.
  • compositions according to the present invention may be administered parenterally in the form of an injectable formulation.
  • Formulations suitable for injection may be formulated using a sterile solvent or any pharmaceutically acceptable carrier.
  • the pharmaceutically acceptable carrier may be selected from sterile water, saline, or the existing cell culture media within the scope of state of the art, but is not limited thereto.
  • Example 1 Materials and Methods Used in the Experimental Procedures
  • pancreatic islet-derived mesenchymal stem cells used in this study were isolated from adult (2.5-3 months of age] Wistar albino rats and characterization studies were completed. All cell culture studies were carried out in Istanbul University, Faculty of Sciences, Department of Biology, Department of Molecular Biology, Primer and Stem Cell Research Laboratory. PI-MSCs between 6 and 9 passages were used in this study.
  • MEM Minimum Essential Medium
  • FBS heat-inactivated fetal bovine serum
  • an antibiotic mixture 100 units/ml penicillin, 100 ⁇ g/ml streptomycin, Gibco-15240-122
  • glutamin (2mM, Gibco, 25030] glutamin 2mM, Gibco, 25030]
  • complete medium at 37°C, 5% C02 and 95% air at a humidified incubator, known as standard culture conditions.
  • Cells reproduced in sterile culture dishes were passaged after filling at least 80% of the culture dish surface. During this process, the cells were allowed to remove from the culture dish by applying 0,25% trypsin-EDTA (Sigma-T4174]. After the trypsin was inactivated by adding complete medium, the cells were collected by centrifuging at 1500 rpm for 5 minutes, and a plating process was performed in a new culture dish after cell counting.
  • PI-MSCs Four different experimental conditions were applied to PI-MSCs. These are i] a complete medium which is a normal growing and developing medium, ii] a complete medium containing 25 mM of glucose, iii] a complete medium containing 1 mM of Valproic acid, and iv] a complete medium containing 1 mM of Valproic acid+25 mM of glucose.
  • D-(+]-Glucose Sigma G7021] was preferred to create glucose-containing conditions and sodium valproate (Sanofi Aventis, Depakine] was used for valproic acid-containing conditions.
  • MTT (3-(4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide] test is a toxicity test aimed at directly assessing cell viability and indirectly cell death.
  • the basis of this method is the reduction of the tetrazolium ring of MTT to a blue-violet, water-insoluble formazan by succinate dehydrogenase, a mitochondrial enzyme, in viable cells.
  • the capability of the cells to reduce MTT is accepted as a criterion of cell viability and formazan density obtained as a result of this reaction is directly proportional to the number of viable cell.
  • Viable cells, the mitochondrial function of which is undistorted, are stained in purple color, whereas dead cells or cells having impaired mitochondrial function are not stained.
  • DMSO dimethylsulfoxide
  • PI-MSC PI-MSC
  • PI-MSC PI-MSC
  • VPA 0.75 mM, 1 mM, 1.5 mM, 2 mM, 2.5 mM, 3 mM, selected in accordance with the literature was applied for 24 hours to determine non-toxic doses of VPA. (Haumaitre et al., 2008].
  • Cells grown in a complete medium were used as a control.
  • 50 mg of MTT (Applichem, A2231 0001] were sterilized by passing through a filter of 0.22 ⁇ after bringing to 10 ml with MEM and dissolving.
  • RNA level was measured in a Qubit Fluorimeter (Invitrogen] using the Qubit® RNA HS Assay Kit (Thermofisher, Q32852].
  • RNA was converted to cDNA by reverse transcriptase (NEB, ProtoScript First Strand cDNA Synthesis Kit, E6300S].
  • NEB reverse transcriptase
  • RNA denaturation was performed at 70°C for 5 minutes.
  • incubation was carried out for 1 hour at 42°C followed by incubation at 80°C for 5 minutes in order to stop the enzyme activity.
  • Thermal Cycler device was used for these steps.
  • Total cDNA level was measured in a Qubit Fluorimeter (Invitrogen] using the Qubit ssDNA Assay Kit.
  • the ability of these cells to release insulin in response to increase in glucose was measured.
  • the cells were incubated in DMEM medium containing 5.5 mM/L of glucose and 0.5% BSA for two hours subsequent to washing with PBS. After the culture medium was collected, the cells were washed with PBS and incubated for two hours in DMEM medium containing glucose (25 mM/L) at high concentration. Insulin levels of the collected media were determined by sandwich ELISA technique and the protocol provided by the kit (Millipore, EZRMI- 13K]. The total protein content of the cells was calculated by the Bradford test and a normalization process was performed proportioning the insulin levels to the protein levels.
  • DCFDA 2',7'-dichlorofluorescein diacetate
  • ROS reactive oxygen species
  • ROS level changes between experimental groups were determined by flow cytometry using DCFDA. Analyses were performed with 50,000 cells from each group and repeated at least 3 times. DCFDA (Sigma-D6883] was added to the cells at a concentration of 10 ⁇ , incubated for 30 min in the dark at standard culture conditions and analyzed on FLl channel on a flow cytometer (BD, FACS Calibur]. The mean fluorescence intensity from the control group was taken as 1 and the ROS levels in the other groups were calculated as fold increase relative to the control group.
  • Example 5 Western Blot The lysis buffer (Invitrogen, FNN0011], to which the protease-phosphatase inhibitor mixture (Serva-39055] was added, was introduced to the cell pellet. This was allowed to stand for 30 minutes with stirring once every 10 minutes on ice. Ultrasonication was performed 5 times for 10 seconds. After centrifugation at 13000 rpm at °C for 10 minutes, the supernatant was stored in a freezer at -86°C for use as a cytoplasmic fraction.
  • lysis buffer Invitrogen, FNN0011]
  • the protease-phosphatase inhibitor mixture Serva-39055
  • the lysis buffer (Invitrogen, FNN0011], to which the protease-phosphatase inhibitor mixture (Serva-39055] was added, was introduced to the cell pellet. This was allowed to stand for 30 minutes with stirring once every 10 minutes on ice. Ultrasonication was performed 5 times for 10 seconds. After centrifugation at 13000 rpm at 4°C for 10 minutes, the supernatant was stored in a freezer at -86°C for use as a cytoplasmic fraction.
  • Samples containing 30 ⁇ g protein for the cytoplasmic fraction and 10 ⁇ g protein for the nuclear fraction were prepared in a volume of 10 ⁇ , and denaturation process was carried out by leaving a total of 20 ⁇ of the mixture obtained by adding 10 ⁇ of laemmli buffer, in a dry block heater heated to 100°C (Techne Dri-Block DB-2D] for 5 minutes. After a brief centrifugation, the wells of the prepared gels were loaded. 25 mM Tris, 192 mM Glisin, 0,1% SDS; pH:8,3 were used a the running buffer. Proteins were run at 70 V for 30 minutes, 100 V for 45 minutes, and at 120 V and +4°C until the running process was terminated. D) Transfer of Proteins from Gel to a Membrane
  • Proteins separated by gel electrophoresis were transferred from the gel to a PVDF (Polyvinylidene fluoride] membrane by wet transfer.
  • PVDF Polyvinylidene fluoride
  • Membranes (Millipore-Immobilon-PSQ] with a pore diameter of 0.2 ⁇ were used for the transfer of proteins less than 20 kDa, while membranes with a pore diameter of 0.45 ⁇ (Millipore-Immobilon-P] were preferred for the transfer of proteins larger than 20 kDa. Transfer was carried out in Tris-Glycine SDS buffer containing 20% methanol for 12 hours at a current intensity of 80mA and + 4°C.
  • Ponceau S staining solution was applied to the membrane for 5 min, and the efficiency of the transfer was checked by observing the band densities. After the bands were observed, 0.1 M NaOH was applied for 5 min to remove the Poncaue S stain from the membrane.
  • the membrane was incubated with a blocking solution prepared with 5% skimmed milk powder for 1 hour at room temperature in a shaker. Membranes were incubated overnight at +4°C with primer antibodies (Table 3.2] prepared with appropriate dilution rate after blocking. After this process, the membranes washed with the washing buffer TBS-T (95% TBS, 5% Twin-20] were treated with the secondary antibodies specific for HRP (horseradish peroxidase]-labeled primer antibody (Table 3.2] for 1 hour at room temperature.
  • HRP horseradish peroxidase
  • miRNAs associated with pluripotency, early pancreatic development, end term pancreas maturation, Type 2 diabetes mellitus and HDACs have been determined.
  • U6 was used as an internal control. Results were calculated in 2 ACT values.
  • 700 ul of this mixture was transferred to 2 ml of collection tubes with filters. Centrifugation was performed at 8000xg for 15 seconds. 700 ul of RWT buffer was added to the columns. Centrifugation was performed at 8000xg at room temperature for 15 seconds. 500 ⁇ of RPE buffer was added and centrifuged at 8000xg at room temperature for 15 seconds. Again, 500 ⁇ of RPE buffer was added and centrifuged at 8000xg at room temperature for 2 minutes. The column was placed in eppendorf tubes of 1.5 ml. 50 ⁇ of Rnase-free water was added to the column. The tube was centrifuged at 8000xg for 1 minute after sealing. miRNAs were allowed to be transferred to the tube together with water.
  • the Qubit® RNA HS Assay Kit (Thermofisher, Q32852] was used to quantify the amount of RNAs obtained as a result of isolation. Tubes of 0.5 ml were prepared for the standards and samples. Solution required for the samples and standards being studied by diluting Qubit RNA HS Reagent in Qubit RNA HS Buffer at a ratio of 1 :200. 190 ⁇ and 199 ⁇ of this solution were transferred to the standard tubes and the sample tubes, respectively. RNA levels were determined by Qubit Fluorimeter 2.0 (Invitrogen] after 10 ⁇ of standards and 1 ⁇ of samples were added, followed by incubation at room temperature for 5 min in the dark.
  • RNA was converted to cDNA using reverse transcriptase enzyme (Qiagene, 2180 3]. For each sample, a total of 20 ⁇ of mixture was formed. This mixture was composed of 4 ⁇ of 5x miScript buffer, 2 ⁇ of lOx miScript mix, 2 ⁇ of miScript Reverse Transcriptase, 5.75 ⁇ of RNase-free water, and 6.25 ⁇ of RNA.
  • incubation was carried out at 95°C for 5 minutes to inactivate the reverse transcriptase enzyme after incubation at 37°C for 60 minutes. Thermal Cycler device (Techne] was used for these steps.
  • reaction mixture prepared in a total volume of 20 ⁇ was composed of 4.5 ⁇ of RNase-free water (QIAGEN], 10 ⁇ of SYBR green (2xQuaintiTect SYBR Green PCR], 2 ⁇ of master mix (lOx miScript Universal Primer], 2 ⁇ of primer mix QIAGEN, miscript primer Assays] and 1.5 ⁇ of cDNA.
  • a reaction mixture of 1.5 ⁇ without Rnase-free water was used as a negative control instead of cDNA.
  • U6 miRNA was used as an internal control.
  • qRT-PCR analysis was performed using the Rotor-Gene Q Series Software 2.3.1 (Qiagen] program.
  • Valproic acid was applied to PI-MSCs at a dose of 0.75-3 mM for 24 hours.
  • the effect of valproic acid on cell viability at the specified dose range was determined by MTT technique and the results are shown in Figure 1.
  • Cell viability values are as follows: 100 ⁇ 0.00% in the control group; 131.40 ⁇ 1.53 in the group treated with 0.75 mM of VP A; 107.50 ⁇ 1.76 in the group treated with 1 mM of VP A; 91.58 ⁇ 2.52 in the group treated with 1.5 mM of VP A; 76.48 ⁇ 2.12 in the group treated with 2 mM of VP A; 58.28 ⁇ 2.08 in the group treated with 2.5 mM of VP A; and 51.63 ⁇ 2.17 in the group treated with 3 mM of VPA. There was found a significant increase in the number of the cells only in the group treated with 0.75 mM of VPA versus control group.
  • VPA concentrations increased after the group treated with 1 mM of VPA. From these results, it was decided to continue with ⁇ -cell differentiation studies by applying 0.75 mM and 1 mM of VPA that are doses increasing or decreasing the cell viability.
  • the insulin + cell ratios recorded after administration of complete media, 25 mM Glucose, 1 mM VPA, and 1 mM VPA + 25 mM Glucose to PI-MSCs for 10, 20 and 30 days are shown in Figure 2.
  • a significant ⁇ -cell differentiation was observed for the group receiving only 1 mM VPA.
  • a significant ⁇ -cell differentiation was observed for the groups receiving 25 mM Glucose, 1 mM VPA and 1 mM VPA+25mM Glucose. It was found that the highest ⁇ -cell differentiation occurred in the groups for which the application was performed for 20 days. From these results, it was decided for the study to be continued with a 20-day experiment period.
  • Figures 4 and 5 show photographs taken at the end of 20 days on the inverted light microscope.
  • the morphological similarity between the glucose-treated PI-MSCs and the complete medium is remarkable.
  • the common characteristic of both groups is the formation of stem cell colonies of largely varying sizes ( Figure 4].
  • This image corresponds to the characteristic of pluripotent cell.
  • the presence of round cells concentrated in colony-like structures in the PI-MSCs treated with VPA and VPA + glucose is noteworthy. It has been observed that the PI-MSCs surrounding the colonies of such cells have decreased in number such that the surrounding of the colonies is emptied and generally, the round cells separated from these colonies are spread to these spaces.
  • ROS reactive oxygen species
  • insulin release was as follows: 0.15 ⁇ 0.01 ng ⁇ g protein in the control group; 0.29 ⁇ 0.05 ng ⁇ g protein in VPA-treated group; and 0.28 ⁇ 0.01 ng ⁇ g protein in VPA+glucose-treated group.
  • the groups were compared with each other, a significantly increased level of insulin secretion was observed in the VPA and VPA + glucose- treated groups compared to the control group (p ⁇ 0,01].
  • insulin release was as follows: 0.15 ⁇ 0.01 ng ⁇ g protein in the control group; 0.28 ⁇ 0.01 ng ⁇ g protein in the glucose group; 0.28 ⁇ 0.01 ng ⁇ g protein in VPA-treated group; and 0.30 ⁇ 0.01 ng ⁇ g protein in VPA+glucose-treated group.
  • the groups were compared with each other, a significantly increased level of insulin secretion was observed in all groups compared to the control group (p ⁇ 0,01 for G and VPA; p ⁇ 0,001 for VPA+glucose].
  • Example 9 Results for Changes in Pluripotent Character
  • Amount of 0CT3/4 in the cytoplasm was found to be 1.17 ⁇ 0.06 for the control group; 0.69 ⁇ 0.18 for glucose-treated group; 0.32 ⁇ 0.10 for VPA-treated group; and 0.37 ⁇ 0.09 for VPA+G-treated group.
  • Nuclear 0CT3/4 ratios were found to be 0.91 ⁇ 0.16 for the control group; 0.45 ⁇ 0.078 for glucose-treated group; 0.34 ⁇ 0.07 for VPA-treated group; and 0.37 ⁇ 0.02 or VPA+G-treated group.
  • Amount of cytoplasmic c-Myc were calculated as 1.07 ⁇ 0.04 for the control group; 0.60 ⁇ 0.015 for glucose-treated group; 0.36 ⁇ 0.06 for VPA-treated group; and 0.52 ⁇ 0.06 for VPA+G-treated group. Ratios of nuclear c-Myc were found to be 0.30 ⁇ 0.06 for the control group; 0.48 ⁇ 0.09 for glucose- treated group; 0.11 ⁇ 0.07 for VPA-treated group; and 0.07 ⁇ 0.01 for VPA+G-treated group. When cytoplasmic c-myc levels were compared, a significant decrease was found in all groups compared to the control group (p ⁇ 0,001].
  • the amount of cytoplasmic Nanog was 1.32 ⁇ 0.21 for the control group; 0.74 ⁇ 0.07 for glucose- treated group; 0.1 ⁇ 0.01 for VPA-treated group; and 0.12 ⁇ 0.01 for VPA + G-treated group.
  • Ratios of nuclear Nanog were 1.27 ⁇ 0.24 for the control group; 0.76 ⁇ 0.07 for glucose-treated group; 0.19 ⁇ 0.01 for VPA-treated group; and 0.13 ⁇ 0.01 for VPA + G-treated group.
  • PI- MSCs Complete medium, 1 mM VP A, 25 mM Glucose and 1 mM VPA+25 mM Glucose was applied to PI- MSCs for 20 days, with the media being changed daily. Subsequently, cytoplasmic and nuclear fractions were obtained from cell lysates and levels of HDACl and 3, H3K9-Ace, H3K9-Met, H4K12-Ace were determined by western blot method. Optical density values of the protein bands were normalized by proportioning to the ⁇ -actin density for the cytoplasmic fraction and to the TF-IIB density for the nuclear fraction.
  • Figure 9 shows the changes in cytoplasmic and nuclear fraction levels of HDACl and 3 among Class I HDACs. Ratios of cytoplasmic HDACl were 0.14 ⁇ 0.01 for the control group; 0.12 ⁇ 0.02 for glucose-treated group; 0.04 ⁇ 0.01 for VPA-treated group; and 0.03 ⁇ 0.01 for VPA + G-treated group. When the results were compared, a significant decrease was found in VPA- and VPA + G- treated groups compared to the control group (p ⁇ 0,001]. When VPA- and VPA + G-treated groups were compared to glucose-treated group, a significant decrease was detected (p ⁇ 0,001].
  • Ratios of nuclear HDACl were 0.43 ⁇ 0.12 for the control group; 0.31 ⁇ 0.07 for glucose-treated group; 0.09 ⁇ 0.05 for VPA-treated group; and 0.07 ⁇ 0.02 for VPA + G-treated group.
  • a significant decrease was detected in VPA- and VPA + G-treated groups compared to the control group (p ⁇ 0,001].
  • VPA- and VPA + G-treated groups were compared to glucose- treated group, a significant decrease was found (p ⁇ 0,01].
  • Ratios of cytoplasmic HDAC3 were 0.15 ⁇ 0.03 for the control group0.12 ⁇ 0.02for glucose-treated group; 0.05 ⁇ 0.01 for VPA-treated group; and 0.04 ⁇ 0.00 for VPA + G-treated group.
  • a significant decrease was observed in VPA- and VPA+G-treated groups compared to glucose-treated group (p ⁇ 0,001].
  • VPA-treated group was compared to glucose-treated group, a significant decrease was found (p ⁇ 0,01].
  • VPA+G-treated group was compared to glucose-treated group, a significant decrease was found (p ⁇ 0,001].
  • Ratios of nuclear HDAC3 were 0.14 ⁇ 0.01 for the control group; 0.11 ⁇ 0.07 for glucose-treated group; 0.03 ⁇ 0.00 for VPA-treated group; and 0.03 ⁇ 0.00 for VPA + G-treated group. A significant decrease was observed in VPA- and VPA+G-treated groups compared to the control group (p ⁇ 0,01]. When VPA- and VPA+G-treated groups were compared to glucose-treated group, a significant decrease was observed (p ⁇ 0,01].
  • Figure 10 shows the results of histone modifications. H4K12-Ace ratios were calculated as 0.006 ⁇ 0.001 in the control group; 0.005 ⁇ 0.001 in VPA-treated group; and 0.075 ⁇ 0.019 in VPA+G- treated group.
  • VPA- treated group compared to the control and glucose-treated group (p ⁇ 0,01).
  • VPA+G-treated group was compared to the control and glucose-treated groups, a significant increase was observed (p ⁇ 0,05). No significant difference was found between VPA- treated group and VPA+G- treated group (p>0,05).
  • H3K9-Met levels were calculated as 0.225 ⁇ 0.001 in the control group, 0.160 ⁇ 0.283 in glucose- treated group, 0.070 ⁇ 0.001 in VPA-treated group; and 0.080 ⁇ 0.042 in VPA + G-treated group. A significant decrease was observed only in VPA- and VPA+G-treated groups compared to the control group (p ⁇ 0,05).
  • H3K9-Ace levels were calculated as 0.076 ⁇ 0.004in the control group, 0.108 ⁇ O.OOlin glucose- treated group, 0.171 ⁇ 0.001 in VPA-treated group; and 0.119 ⁇ 0.130 in VPA + G-treated group.
  • a significant increase was observed in VPA-treated group compared to the control and glucose- treated group (p ⁇ 0,001].
  • a significant decrease (p ⁇ 0.05] was found when VPA+G-treated group was compared with VPA-treated group, and a significant increase (p ⁇ 0,05] was found when VPA+G-treated group was compared with the control group.

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Abstract

La présente invention concerne l'utilisation d'au moins un microARN choisi dans le groupe constitué de miR -18 a, miR -19 b, miR -30 d, miR -124, miR -146 a, miR -184, miR -435, miR-433-5 p et miR -503, ou d'un ADN, d'un ARNm ou d'un ARNnc associé, dans le diagnostic et le traitement de maladies liées à l'insuline telles que le diabète sucré et l'obésité.
PCT/TR2017/050737 2016-12-30 2017-12-31 Utilisation de certains miarn destinés au diagnostic et au traitement de maladies associées à l'insuline Ceased WO2018125019A2 (fr)

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TR2016/20327 2016-12-30
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CN113122536A (zh) * 2021-03-31 2021-07-16 南通大学 一种促进神经干细胞向神经元分化的长链非编码rna及其筛选方法
CN113774127A (zh) * 2021-09-24 2021-12-10 南京医科大学 血清细胞外囊泡miR-503-5p在制备2型糖尿病发病的诊断试剂盒中的应用
WO2022060178A1 (fr) * 2020-09-21 2022-03-24 고려대학교 산학협력단 Procédé pour la prédiction de la récurrence ou du pronostic de l'ulcère du pied diabétique en utilisant la méthylation spécifique du gène
WO2022065859A1 (fr) * 2020-09-24 2022-03-31 성균관대학교산학협력단 Procédé de reprogrammation directe de cellules somatiques en cellules bêta pancréatiques à l'aide de microarn, et composition de différenciation
CN114555070A (zh) * 2019-08-26 2022-05-27 经期药丸私人有限责任公司 月经周期诱发的症状治疗
KR20240108335A (ko) * 2020-09-21 2024-07-09 고려대학교 산학협력단 유전자의 특이적 메틸화를 이용하여 당뇨족부궤양 재발 또는 예후를 예측하는 방법
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WO2014135653A1 (fr) * 2013-03-06 2014-09-12 Royal College Of Surgeons In Ireland Diagnostic et traitement de troubles du métabolisme
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CN114555070A (zh) * 2019-08-26 2022-05-27 经期药丸私人有限责任公司 月经周期诱发的症状治疗
WO2022060178A1 (fr) * 2020-09-21 2022-03-24 고려대학교 산학협력단 Procédé pour la prédiction de la récurrence ou du pronostic de l'ulcère du pied diabétique en utilisant la méthylation spécifique du gène
KR20240108335A (ko) * 2020-09-21 2024-07-09 고려대학교 산학협력단 유전자의 특이적 메틸화를 이용하여 당뇨족부궤양 재발 또는 예후를 예측하는 방법
KR102850036B1 (ko) 2020-09-21 2025-08-26 고려대학교 산학협력단 유전자의 특이적 메틸화를 이용하여 당뇨족부궤양 재발 또는 예후를 예측하는 방법
WO2022065859A1 (fr) * 2020-09-24 2022-03-31 성균관대학교산학협력단 Procédé de reprogrammation directe de cellules somatiques en cellules bêta pancréatiques à l'aide de microarn, et composition de différenciation
CN113122536A (zh) * 2021-03-31 2021-07-16 南通大学 一种促进神经干细胞向神经元分化的长链非编码rna及其筛选方法
CN113122536B (zh) * 2021-03-31 2023-06-20 南通大学 一种促进神经干细胞向神经元分化的长链非编码rna及其筛选方法
CN113774127A (zh) * 2021-09-24 2021-12-10 南京医科大学 血清细胞外囊泡miR-503-5p在制备2型糖尿病发病的诊断试剂盒中的应用
CN113774127B (zh) * 2021-09-24 2022-06-24 南京医科大学 血清细胞外囊泡miR-503-5p在制备2型糖尿病发病的诊断试剂盒中的应用
US12485100B2 (en) 2023-01-06 2025-12-02 Period Pill Bv Treatment of menstrual cycle-induced symptoms

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