Cardiac fibroblast-derived extracellular matrix and injectable formulations thereof for treatment of ischemic disease or injury

Patent metadata

Field	Value
Publication number	US-2016354447-A1
Application number	US-201615173367-A
Country	US
Kind code	A1
Filing date	Jun 3, 2016
Priority date	Jun 3, 2015
Publication date	Dec 8, 2016
Grant date	—

Abstract

Compositions and methods using an engineered cardiac fibroblast-derived 3-dimensional extracellular matrix (ECM) are disclosed. The ECM includes the structural proteins fibronectin, collagen type I, collagen type III, and elastin, and from 60% to 90% of the structural proteins present in the engineered extracellular matrix are fibronectin. The compositions, which can be used to treat cardiac disease or ischemic disease or injury, are injectable compositions, where the ECM is diced into small fragments or lyophilized into a powder. The disclosed methods include a method of treating ischemic disease or injury by contacting a cell free patch made from the ECM with the injured tissue, without the concomitant delivery of therapeutic cells, and a method of treating ischemic limb injury by contacting a patch, either by itself or seeded with therapeutic cells, with the injured limb tissue.

First claim

We claim:

1 . An injectable composition for treating cardiac disease, cardiac injury, or ischemic injury, comprising:
(a) one or more fragments of an engineered cardiac fibroblast cell-derived extracellular matrix comprising the structural proteins fibronectin, collagen type I, collagen type III, and elastin, wherein from 60% to 90% of the structural proteins present in the engineered extracellular matrix are fibronectin, wherein the one or more fragments are small enough to pass through the injection opening of an 18 gauge hypodermic needle; and (b) an injectable pharmaceutically acceptable carrier.

2 . The composition of claim 1 , wherein the structural proteins of the engineered cardiac extracellular matrix are not chemically cross-linked.

3 . The composition of claim 1 , wherein the one or more fragments have a thickness of 20-500 μm.

4 . The composition of claim 1 , wherein the one or more fragments are in the form of a lyophilized powder.

5 . The composition of claim 1 , wherein the engineered cardiac extracellular matrix further comprises one or more of the matricellular proteins latent transforming growth factor beta 1 (LTGFB-1), latent transforming growth factor beta 2 (LTGFB-2), connective tissue growth factor (CTGF), secreted protein acidic and rich in cysteine (SPARC), versican core protein (VCAN), galectin 1, galectin 3, matrix gla protein (MGP), sulfated glycoprotein 1, and biglycan.

6 . The composition of claim 1 , wherein the composition is essentially devoid of intact cardiac fibroblast cells.

7 . The composition of claim 6 , wherein the composition is cell free.

8 . The composition of claim 1 , further comprising one or more cells that are therapeutic for cardiac disease, cardiac injury, or ischemic injury.

9 . The composition of claim 8 , wherein the one or more cells that are therapeutic for cardiac disease, cardiac injury, or ischemic injury are selected from the group consisting of skeletal myoblasts, embryonic stem (ES) cells or derivatives thereof, induced pluripotent stem (iPS) cells or derivatives thereof, multipotent adult germline stem cells (maGCSs), bone marrow mesenchymal stem cells (BMSCs), very small embryonic-like stem cells (VSEL cells), endothelial progenitor cells (EPCs), cardiopoietic cells (CPCs), cardiosphere-derived cells (CDCs), multipotent Isl1+ cardiovascular progenitor cells (MICPs), epicardium-derived progenitor cells (EPDCs), adipose-derived stem cells, human mesenchymal stem cells derived from iPS or ES cells, human mesenchymal stromal cells derived from iPS or ES cells, and combinations thereof.

10 . The composition of claim 1 , further comprising one or more exogenous non-cellular therapeutic agents.

11 . The composition of claim, 10 , wherein the one or more exogenous non-cellular therapeutic agents include one or more growth factors.

12 . The composition of claim 11 , wherein the one or more growth factors are selected from the group consisting of epidermal growth factor (EDF), transforming growth factor-α (TGF-α), hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), platelet derived growth factor (PDGF), fibroblast growth factor 1 (FGF-1), fibroblast growth factor 2 (FGF-2), transforming growth factor-β (TGF-β), stromal derived factor-1 (SDF-1), and keratinocyte growth factor (KGF).

13 . The composition of claim 1 wherein the one or more fragments are small enough to pass through the injection opening of a 27 gauge hypodermic needle.

14 . A method for treating a subject having a cardiac disease, cardiac injury, ischemic disease or ischemic injury, comprising injecting into the subject an effective amount of the composition of claim 1 , whereby the severity of the cardiac disease, cardiac injury, ischemic disease or ischemic injury is decreased.

15 . The method of claim 14 , wherein the cardiac disease, cardiac injury, ischemic disease or ischemic injury is caused by an acute myocardial infarct, heart failure, viral infection, bacterial infection, congenital defect, stroke, diabetic foot ulcer, peripheral artery disease (PAD), PAD-associated ulcer, or limb ischemia.

16 . A method for treating a subject having an ischemic disease or injury, comprising contacting a tissue of the subject that is exhibiting an ischemic injury with a cell free engineered cardiac fibroblast cell-derived extracellular matrix having a thickness of 20-500 μm comprising the structural proteins fibronectin, collagen type I, collagen type III, and elastin, wherein from 60% to 90% of the structural proteins present in the engineered extracellular matrix are fibronectin, and wherein no cells are seeded onto the engineered cardiac fibroblast cell-derived extracellular matrix,
whereby the severity of the ischemic disease or injury is decreased.

17 . The method of claim 16 , wherein the cell free engineered cardiac extracellular matrix further comprises one or more of the matricellular proteins latent transforming growth factor beta 1 (LTGFB-1), latent transforming growth factor beta 2 (LTGFB-2), connective tissue growth factor (CTGF), secreted protein acidic and rich in cysteine (SPARC), versican core protein (VCAN), galectin 1, galectin 3, matrix gla protein (MGP), sulfated glycoprotein 1, and biglycan.

18 . The method of claim 16 , further comprising contacting the tissue of the subject that is exhibiting an ischemic disease or injury with one or more exogenous non-cellular therapeutic agents.

19 . The method of claim 18 , wherein the one or more non-cellular therapeutic agents include one or more growth factors.

20 . The method of claim 19 , wherein the one or more growth factors are selected from the group consisting of stromal derived factor-1 (SDF-1), epidermal growth factor (EDF), transforming growth factor-α (TGF-α), hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), platelet derived growth factor (PDGF), fibroblast growth factor 1 (FGF-1), fibroblast growth factor 2 (FGF-2), transforming growth factor-β (TGF-β), and keratinocyte growth factor (KGF).

21 . The method of claim 16 , wherein the ischemic disease or injury is caused by myocardial infarct, stroke, diabetic foot ulcer, peripheral artery disease (PAD), PAD-associated ulcer, or limb ischemia.

22 . The method of claim 21 , wherein the limb ischemia is the result of atherosclerosis or diabetes.

23 . A method for treating a subject having an ischemic limb injury, comprising contacting a tissue of the subject that is exhibiting an ischemic limb injury with an engineered cardiac fibroblast cell-derived extracellular matrix having a thickness of 20-500 μm comprising the structural proteins fibronectin, collagen type I, collagen type III, and elastin, wherein from 60% to 90% of the structural proteins present in the engineered extracellular matrix are fibronectin,
whereby the severity of the ischemic limb injury is decreased.

24 . The method of claim 23 , wherein the engineered cardiac extracellular matrix further comprises one or more of the matricellular proteins latent transforming growth factor beta 1 (LTGFB-1), latent transforming growth factor beta 2 (LTGFB-2), connective tissue growth factor (CTGF), secreted protein acidic and rich in cysteine (SPARC), versican core protein (VCAN), galectin 1, galectin 3, matrix gla protein (MGP), sulfated glycoprotein 1, and biglycan.

25 . The method of claim 23 , wherein the engineered cardiac extracellular matrix is

Assignees

• Wisconsin Alumni Res Found

Inventors

• Schmuck Eric
• Raval Amish

Classifications

• A61P9/04

  Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure · CPC title

• A61P3/10

  for hyperglycaemia, e.g. antidiabetics · CPC title

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• A61P31/12

  Antivirals · CPC title

• A61P9/10

  for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis · CPC title