EP-3746179-B1 - METHODS AND COMPOSITIONS FOR TREATING AND PREVENTING DIABETES

Inventors

• LEVENBERG, SHULAMIT
• KARNIELI, EDDY
• LEWIS, Eli Chaim
• BECKERMAN, Margarita
• HAREL, Chava

Dates

Publication Date

20260513

Application Date

20190204

Claims (9)

1. A scaffold-cell construct comprising: a. a porous scaffold comprising at least one biocompatible polymer comprising a combination of poly-1-lactic acid (PLLA) and polylactic glycolic acid (PLGA), or collagen; and b. a cell population deposited on or in said scaffold comprising recombinant myoblasts: (i) comprising an exogenous polynucleotide encoding glucose transporter type 4 (GLUT4); (ii) over-expressing said GLUT4; and (iii) having increased GLUT4 activity, compared to control wild type myoblast cells, and wherein said scaffold-cell construct is transplantable.
2. The scaffold-cell construct of claim 1, being characterized by having greater glucose uptake rates compared to said control wild type myoblast cells.
3. The scaffold-cell construct of claim 1 or 2, wherein said PLLA and said PLGA are in a ratio of 3:1 - 1:3 w/w ratio.
4. The scaffold-cell construct of any one of claims 1-3, wherein said biocompatible polymer comprises interconnected pores, wherein at least 80% of said pores have a diameter of between 200 and 600 microns.
5. The scaffold-cell construct of any one of claims 1-4, for use in restoring glucose homeostasis levels in a subject in need thereof, wherein: a. said glucose levels are: i. less than or equal to 100 mg/dL at fasting; and ii. lower than 140 mg/dL postprandial; or b. said glucose levels are: i. less than or equal to 120 mg/dL at fasting; and ii. lower than 160 mg/dL postprandial.
6. The scaffold-cell construct of any one of claims 1-5, for use in the treatment or prevention of diabetes mellitus or metabolic syndrome in a subject in need thereof, preferably wherein said metabolic syndrome is selected from: obesity, pre-diabetes and insulin resistance or related to insulin resistance.
7. A composition comprising the scaffold-cell construct of any one of claims 1-4.
8. A method of making a scaffold-cell construct for restoring glucose homeostatic levels in a diabetic subject in need thereof, the method comprising contacting recombinant myoblasts: (i) comprising an exogenous polynucleotide encoding GLUT4; (ii) over-expressing said GLUT4; and (iii) having increased GLUT4 activity, compared to control wild type myoblast cells, with a scaffold comprising at least one biocompatible polymer comprising a combination of PLLA and PLGA or collagen, and wherein said scaffold-cell construct is transplantable.
9. The method of claim 8, further comprising culturing said recombinant myoblasts on or in said scaffold for at least 7 days.

Description

FIELD OF INVENTION The present invention is in the field of biomedical engineering. BACKGROUND OF THE INVENTION Diabetes mellitus (DM) is a disease that occurs in all populations and age groups and affects more than 10% of the Western world. It is the sixth leading cause of death in the United States, affecting between 6% and 7% of the US population equating to about 16 million people. The two most common general categories of this disease are termed type 1 diabetes (DM1) and type 2 diabetes (DM2). The number of people with diabetes is expected to reach 300 million in 2030, of which 90% will be DM2. Obesity is a major environmental factor contributing to the increasing incidence of DM2. Modern lifestyle, high-fat diet and lack of exercise were shown to trigger the development of DM2 in overweight patients with impaired glucose tolerance, while increased levels of markers and mediators of inflammation and oxidative stress components correlated with impaired insulin action (insulin resistance). DM2 is a heterogeneous, polygenic disorder characterized by defects in insulin action in tissues (insulin resistance) and/or defects in pancreatic insulin secretion (beta cell dysfunction), which eventually results in loss of pancreatic insulin-secreting (beta) cells. The associated complications of diabetes are cardiovascular disease, peripheral vascular disease, stroke, diabetic neuropathy, diabetic nephropathy and diabetic retinopathy. These result in increasing disability, reduced life expectancy and enormous health costs. The development of these diabetes-related complications can be significantly reduced, partially prevented and retarded with control of blood glucose levels as close to normal as possible. However, in spite of the current knowledge and new treatment protocols, many patients currently do not reach the desired treatment goals. DM2 is a progressive and complex disorder that is difficult to treat effectively in the long term. The treatment begins with a well-balanced diet combined with exercise. Unfortunately, the majority of patients are unable to achieve or sustain near normo-glycemia without oral antidiabetic agents; a sizeable proportion of patients will eventually require oral and/or injectable hypoglycemic drugs and/or insulin therapy to maintain long-term glycemic control. The frequent need for escalating therapy reflects progressive loss of islet beta-cell function, usually in the presence of obesity-related insulin resistance. Insulin resistance is a key component in the pathogenesis of DM2. It is a state of resistance to the action of insulin in its target tissues, i.e., impaired insulin stimulation of glucose transport in adipose tissue and skeletal muscle, and reduced inhibition of glucose production and release in the liver. One of the earliest defects detected in DM2 is reduction of cellular content and an impaired function of the insulin-responsive glucose transporter type 4, a member of the glucose transport proteins (GLUTs) mediating glucose uptake in eukaryotic cells. GLUT4 is the main glucose transporter regulating glucose entry from the blood into adipose and muscle tissues upon insulin stimulation. Fussenegger and Xie (EP Patent No. 3272877) describe methods for producing β-cell-mimetic cells as well as methods of using the β-cell-mimetic cells as a medicament and methods of using β-cell-mimetic cells for the prevention, delay of progression or treatment of a metabolic disease in a subject. Levenberg and Francis (International Patent Application No. PCT/IL2008/000337, published as WO2008111064) describe a heterogeneous population of cells seeded on a surface of a scaffold, wherein the heterogeneous population of cells includes at least one pancreatic islet, endothelial cells and fibroblast cells, as well as methods of generating same and uses thereof. Atkinson et al., 2013 describe that moderate GLUT4 overexpression improves insulin sensitivity and fasting triglyceridemia in high-fat diet-fed transgenic mice. Otaegui et al., 2002 describe that glucose-regulated glucose uptake by transplanted muscle cells expressing glucokinase counteracts diabetic hyperglycemia. Michael et al., 2001 describe the restoration of insulin-sensitive glucose transporter (GLUT4) gene expression in muscle cells by the transcriptional coactivator PGC-1. SUMMARY OF THE INVENTION According to the first aspect, there is provided a scaffold-cell construct comprising: (a) a porous scaffold comprising at least one biocompatible polymer comprising a combination of poly-1-lactic acid (PLLA) and polylactic glycolic acid (PLGA), or collagen; and (b) a cell population deposited on or in the scaffold comprising recombinant myoblasts: (i) comprising an exogenous polynucleotide encoding glucose transporter type 4 (GLUT4); (ii) over-expressing the GLUT4; and (iii) having increased GLUT4 activity, compared to control wild type myoblast cells, and wherein the scaffold-cell construct is transplantable. According to another