Prevention of progressive heart failure

Abstract

To provide methods for treating or preventing progressive heart failure.SOLUTION: A method for treating or preventing progressive heart failure in a myocardial infarction subject, comprises administering to the subject a population of mesenchymal lineage precursor or stem cells and / or progeny thereof and / or soluble factors derived therefrom, where the subject has a proximal left anterior descending (LAD) lesion.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present disclosure relates to a method for preventing progressive heart failure in a subject with persistent left ventricular (LV) dysfunction. Such a method may be used to treat or prevent progressive heart failure in a subject with proximal left anterior descending artery (LAD) lesions and persistent left ventricular dysfunction. [Background technology]

[0002] Myocardial infarction (MI) remains one of the leading causes of mortality and morbidity in developed countries. A recent update of U.S. Medicare records was published evaluating data including 350,509 acute MI hospitalizations in patients over 65 years of age who were discharged alive after the event (Schuster et al. (2004) Physiol Heart Circa Physiol., 287(2):525-32). Within the first year after the index event, 25.9% of MI patients died, and 50.5% were rehospitalized. In the month following the MI, the likelihood of death was 21 times higher than in the general Medicare age population, and the likelihood of hospitalization was 12 times higher.

[0003] Patients with larger infarcts after MI and patients with more post-infarction LV dysfunction are at significantly increased risk of experiencing mid- to long-term cardiac events and death. Specifically, subjects with anterior infarcts, larger infarcts, and more LV dysfunction in the post-infarction period are at significantly increased risk of experiencing mid- to long-term cardiac events and death (Eitel et al. (2010) J Am Coll Cardiol., 55:2470-9).

[0004] Infarction due to proximal LAD occlusion remains a major risk factor for progressive LV dilation, remodeling, and symptomatic progressive heart failure. In the post-angioplasty stage, 3-year mortality after MI remains higher for proximal LAD lesions (10% vs. 3% for distal LAD) (Elsman et al. (2006) Am J Cardiol., 97(8):1137-41), most likely due to the 40% larger infarct size associated with this anatomical lesion (Elsman et al. (2006) Am J Cardiol., 97(8):1137-41). Since infarct size greater than 18.5% has been prospectively shown to result in a 30% incidence of heart failure-related major adverse cardiac events (HF-MACE, defined as hospitalization or death due to heart failure) over 2 years (Wu et al. (2008) Heart, 94:730-736), this suggests that the population of MI patients with proximal LAD lesions, low ejection fraction, and large infarcts are at highest risk for subsequent HF-MACE.

[0005] Clearly, there is a need in the art for treating or preventing progressive heart failure. Summary of the Invention

[0006] The present disclosure is based on the unexpected identification of a population of myocardial infarction (MI) subjects who would respond well to stem cell therapy. A number of MI subjects with elevated troponin or CK-MB (more than 4 times the upper limit of normal (ULN)), regional cardiac wall abnormalities, and reduced global left ventricular systolic function (less than 45% and more than 20%) as determined by screening cardiac imaging performed within approximately 24 hours of MI were administered stem cell therapy or placebo therapy immediately after MI.

[0007] We found that overall left ventricular systolic function returned to normal levels in most of these subjects approximately 5 days after MI. The administration of stem cell therapy did not provide any therapeutic improvement over placebo therapy in these subjects.

[0008] Surprisingly, the application of stem cell therapy provided a significant therapeutic improvement compared to placebo treatment in subjects with proximal left anterior descending artery (LAD) lesions. These results indicate that stem cell therapy may be useful for treating or preventing progressive heart failure in a subpopulation of MI subjects, particularly those with proximal LAD lesions.

[0009] Thus, in one example, the present disclosure provides a method of treating or preventing progressive heart failure in a subject with myocardial infarction, the method comprising administering a population of mesenchymal progenitor or stem cells and / or their progeny and / or soluble factors derived therefrom to the subject, the subject having a proximal left anterior descending artery (LAD) lesion.

[0010] In another example, the method includes the steps of: i) selecting a subject having a proximal left anterior descending artery (LAD) lesion; and ii) administering to the subject a population of mesenchymal stem or progenitor cells and / or their progeny and / or soluble factors derived therefrom.

[0011] The inventors also confirmed that subjects with proximal LAD lesions who responded well to stem cell therapy also had persistently low ejection fractions approximately 5 days after MI. These results indicate that the methods of the present disclosure may also be useful for treating or preventing progressive heart failure in subjects with proximal LAD lesions and persistently low ejection fractions.

[0012] Thus, in another example, the present disclosure provides a method of treating progressive heart failure in a subject with myocardial infarction, the method comprising administering a population of mesenchymal progenitor or stem cells and / or their progeny and / or soluble factors derived therefrom to the subject, the subject having proximal left anterior descending artery (LAD) lesions and having persistent left ventricular dysfunction.

[0013] In one example, the subject also has an elevated left ventricular end-systolic volume (LVESV) of greater than 70 mL. In one example, the LVESV is greater than 80 mL, greater than 90 mL, greater than 100 mL, greater than 110 mL, or greater than 120 mL. In another example, the LVESV is greater than 80 mL / m 2 More than 90 mL / m 2 More than 100 mL / m 2 exceeding 110 mL / m 2 or more than 120 mL / m 2 Exceeds.

[0014] In some instances, the subject has a left ventricular ejection fraction (LVEF) of less than about 55%. In other instances, the subject has an LVEF of less than about 45%. In other instances, the subject has an LVEF of less than about 40%. In some instances, the LVEF is measured by cardiovascular magnetic resonance imaging (cMR).

[0015] The inventors have also determined that the timing of administration after MI can also be beneficial to the subject. Thus, in one example, mesenchymal precursor cells or stem cells and / or their progeny and / or soluble factors derived therefrom are administered between about 1 and 7 days after myocardial infarction. In one example, mesenchymal precursor cells or stem cells and / or their progeny and / or soluble factors derived therefrom are administered between about 2 and 7 days after myocardial infarction. In another example, mesenchymal precursor cells or stem cells and / or their progeny and / or soluble factors derived therefrom are administered between about 3 and 5 days after myocardial infarction.

[0016] The inventors have further characterized the population of subjects who may benefit from the methods of the present disclosure based on serum biomarker levels compared to the upper limit of normal (ULM). In one example, the subject has creatine kinase-MB and / or troponin greater than about two times the upper limit of normal. In another example, the subject has creatine kinase-MB and / or troponin and / or myoglobin greater than about four times the upper limit of normal.

[0017] The inventors have further characterized a population of subjects who may benefit from the methods of the present disclosure based on infarct size. In some instances, the subject has an infarct size between about 10-25% of the left ventricle. In other instances, the subject has an infarct size greater than about 18.5% of the left ventricle. In some instances, the infarct size is measured by cMR.

[0018] In another example, the methods of the present disclosure include STRO-1 + The method comprises administering a population of mesenchymal precursor or stem cells enriched for cells and / or their progeny and / or soluble factors derived therefrom.

[0019] In another example, the methods of the present disclosure include STRO-1 bright The method comprises administering a population of mesenchymal precursor or stem cells enriched for cells and / or their progeny and / or soluble factors derived therefrom.

[0020] In some instances, the population of mesenchymal progenitor or stem cells express tissue-nonspecific alkaline phosphatase (TNAP), and / or the progeny cells and / or soluble factors are derived from mesenchymal progenitor or stem cells that express TNAP.

[0021] In some instances, the population of mesenchymal progenitor or stem cells is 6 express angiopoietin-1 (Ang1) in an amount of at least 0.1 μg per cell, and / or the progeny cells and / or soluble factors are 6 In one example, the population of mesenchymal progenitor or stem cells is derived from mesenchymal progenitor or stem cells that express Ang1 in an amount of at least 0.1 μg per cell. 6 In some instances, the population of mesenchymal progenitor or stem cells expresses an amount of Ang1 of at least 0.5 μg per cell. 6 In some instances, the population of mesenchymal progenitor or stem cells expresses an amount of Ang1 of at least 0.7 μg per cell. 6 Express Ang1 in amounts of at least 1 μg per cell.

[0022] In some instances, the population of mesenchymal progenitor or stem cells is 6 express vascular endothelial growth factor (VEGF) in an amount of less than about 0.05 μg per cell, and / or the progeny and / or soluble factors are less than 10 6 The population of mesenchymal precursor or stem cells is derived from mesenchymal precursor or stem cells that express VEGF and / or progeny in an amount of less than about 0.05 μg per cell. In one example, the population of mesenchymal precursor or stem cells is derived from mesenchymal precursor or stem cells that express VEGF and / or progeny in an amount of less than about 0.05 μg per cell. 6 They express VEGF in amounts less than about 0.03 μg per cell.

[0023] In some examples, the population of mesenchymal precursor or stem cells expresses a ratio of Ang1:VEGF of at least about 2:1, and / or progeny cells and / or soluble factors are derived from mesenchymal precursor or stem cells that express a ratio of Ang1:VEGF of at least about 2:1. In some examples, the population of mesenchymal precursor or stem cells expresses a ratio of Ang1:VEGF of at least about 10:1. In some examples, the population of mesenchymal precursor or stem cells expresses a ratio of Ang1:VEGF of at least about 20:1. In some examples, the population of mesenchymal precursor or stem cells expresses a ratio of Ang1:VEGF of at least about 30:1.

[0024] In one example, the population of mesenchymal precursor cells or stem cells and / or their progeny and / or soluble factors derived therefrom is administered systemically. In some examples, the population of mesenchymal precursor cells or stem cells and / or their progeny and / or soluble factors derived therefrom is administered intravenously, intramuscularly, or intranasally. For example, the population of mesenchymal precursor cells or stem cells and / or their progeny and / or soluble factors derived therefrom can be administered intravenously.

[0025] In one example, multiple doses of a population of mesenchymal precursor or stem cells and / or their progeny and / or soluble factors derived therefrom are administered.

[0026] In one example, the method of the present disclosure 6 ~8×10 8 In some examples, the methods of the disclosure include administering between 1.2 x 10 cells. 8 ~4×10 8 In some examples, the methods of the present disclosure include administering between about 1.5 x 10 cells. 8 The method includes administering cells.

[0027] In some instances, the population of cells and / or progeny cells are autologous or allogeneic, and / or the soluble factors are derived from autologous or allogeneic cells.

[0028] In some instances, a population of cells and / or their progeny is expanded in culture prior to administration and / or obtaining the soluble factor.

[0029] In some instances, the mesenchymal precursor or stem cells and / or their progeny and / or soluble factors derived therefrom are administered in the form of a composition comprising the mesenchymal precursor or stem cells and / or their progeny and / or soluble factors derived therefrom and a carrier and / or excipient. For example, the composition may include a cryopreservation agent. Thus, in one example, the present disclosure relates to a population of mesenchymal precursor cells or stem cells and / or their progeny and / or soluble factors derived therefrom for use in treating or preventing progressive heart failure in a subject with myocardial infarction (MI), wherein the subject has proximal left anterior descending (LAD) artery disease. In another example, the present disclosure relates to the use of a population of mesenchymal precursor cells or stem cells and / or their progeny and / or soluble factors derived therefrom in the manufacture of a medicament for treating or preventing progressive heart failure in a subject with myocardial infarction (MI), wherein the subject has proximal left anterior descending (LAD) artery disease. In these examples, the subject may have persistent left ventricular dysfunction. For example, the subject may have an LVEF of less than about 55%. In another example, the subject has an LVEF of less than about 45%. In another example, the subject has an LVEF of less than about 40%. In these examples, the mesenchymal precursor cells or stem cells and / or their progeny and / or soluble factors derived therefrom can be administered between about 1 and 7 days after myocardial infarction. For example, the mesenchymal precursor cells or stem cells and / or their progeny and / or soluble factors derived therefrom can be administered between about 3 and 5 days after myocardial infarction. In these examples, the subject can have creatine kinase-MB and / or troponin greater than about two times the upper limit of normal. In another example, the subject can have creatine kinase-MB and / or troponin and / or myoglobin greater than about four times the upper limit of normal. In these examples, the subject can have an infarct size between about 10-25% of the left ventricle. In another example, the subject can have an infarct size greater than about 18.5% of the left ventricle. In these examples, the LVEF or infarct size can be measured by cMR. [Brief explanation of the drawings]

[0030] [Figure 1] Comparison of placebo and remestemcel-L treatment groups at baseline. [Figure 2] Comparison of placebo and remestemcel-L treatment groups for change from baseline at 6 months. [Figure 3] Comparison of placebo and remestemcel-L treatment groups in the % of patients with a change in left ventricular end-diastolic volume less than -10 ml after 6 months. Measured change from baseline. [Figure 4] 1 shows the change in LVESV values ​​at 6 months in subjects receiving placebo (control) and MPC (150 million cells) stratified by LVESV greater than 70 mL. [Figure 5] 1 shows the change in LVESV values ​​at 6 months in subjects receiving placebo (control) and MPC (150 million cells) stratified by LVESV greater than 80 mL. [Figure 6] 1 shows the change in LVESV values ​​at 6 months in subjects receiving placebo (control) and MPC (150 million cells) stratified by LVESV greater than 90 mL. [Figure 7] 1 shows the change in LVESV values ​​at 6 months in subjects receiving placebo (control) and MPC (150 million cells) stratified by LVESV greater than 100 mL. [Figure 8] 1 shows the change in LVESV values ​​at 6 months in subjects receiving placebo (control) and MPC (150 million cells) stratified by LVESV greater than 110 mL. [Figure 9] 1 shows the change in LVESV values ​​at 6 months in subjects receiving placebo (control) and MPC (150 million cells) stratified by LVESV greater than 120 mL.

[0031] General Techniques and Definitions Unless specifically stated otherwise, all technical and scientific terms used herein should be construed to have the same meaning as commonly understood by one of ordinary skill in the art (e.g., in molecular genetics, molecular biology, cell culture, stem cell differentiation, immunology, immunohistochemistry, protein chemistry, and biochemistry).

[0032] Unless otherwise indicated, the stem cell, cell culture, and surgical techniques utilized in this disclosure are standard procedures, well known to those skilled in the art. Such techniques are described and explained throughout the literature in sources such as J. Perbal, A Practical Guide to Molecular Cloning, John Wiley and Sons (1984), J. Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press (1989), T.A. Brown (editor), Essential Molecular Biology: A Practical Approach, Volumes 1 and 2, IRL Press (1991), D.M. Glover and B.D. Hames (editors), and F.M. Ausubel et al. (editors), Current Protocols in Molecular Biology, Greene Pub. Associates and Wiley-Interscience (1988, including all updates until present), Ed Harlow and David Lane (editors), Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, (1988), and J.E. Coligan et al. (editors), Current Protocols in Immunology, John Wiley & Sons (including all updates until present).

[0033] Throughout this specification, unless specifically stated otherwise or the context requires otherwise, references to a single step, composition of matter, group of steps, or group of compositions of matter should be interpreted as encompassing one and more (i.e., one or more) of those steps, compositions of matter, groups of steps, or group of compositions of matter.

[0034] The present disclosure is not to be limited in scope by the specific embodiments described herein, which are for purposes of illustration only. Functionally equivalent products, compositions, and methods are clearly within the scope of the present disclosure as described herein.

[0035] Any example disclosed herein should be construed to apply mutatis mutandis to any other example, unless specifically stated otherwise.

[0036] The term "and / or," e.g., "X and / or Y," should be understood to mean either "X and Y" or "X or Y," and should be interpreted as providing explicit support for both meanings or either meaning.

[0037] As used herein, unless stated to the contrary, the term "about" refers to ±10%, more preferably plus or minus 5%, of a given value.

[0038] Throughout this specification the word "comprise" or variations such as "comprises" or "comprising" will be understood to mean the inclusion of a stated element, integer, or step or group of elements, integers, or steps, but not the exclusion of any other element, integer, or step or group of elements, integers, or steps.

[0039] Mesenchymal progenitor cells As used herein, the term "mesenchymal progenitor or stem cell" refers to an undifferentiated, multipotent cell capable of self-renewal while maintaining pluripotency and the ability to differentiate into multiple cell types, either of mesenchymal origin, e.g., osteoblasts, chondrocytes, adipocytes, stromal cells, fibroblasts, and tendons, or of non-mesodermal origin, e.g., hepatocytes, neurons, and epithelial cells. For the avoidance of doubt, "mesenchymal progenitor cell" refers to a cell that can differentiate into mesenchymal cells, such as bone cells, cartilage cells, and adipocytes, as well as fibrous connective tissue.

[0040] The term "mesenchymal precursor or stem cells" includes both parent cells and their undifferentiated progeny. The term also includes mesenchymal precursor cells, multipotent stromal cells, mesenchymal stem cells (MSCs), perivascular mesenchymal precursor cells, and their undifferentiated progeny.

[0041] Mesenchymal progenitor or stem cells can be autologous, xenogeneic, syngeneic, or isogenic. Autologous cells are isolated from the same individual into whom they are to be reimplanted. Allogeneic cells are isolated from a donor of the same species. Xenogeneic cells are isolated from a donor of another species. Syngeneic or isogenic cells are isolated from genetically identical organisms, such as twins, clones, or highly inbred research animal models.

[0042] Mesenchymal progenitor or stem cells reside primarily in bone marrow, but have also been shown to reside in a wide variety of host tissues, including, for example, umbilical cord blood and cord, adult peripheral blood, adipose tissue, trabecular bone, and dental pulp.

[0043] In one example, the mesenchymal progenitor cells or stem cells are STRO-1+ mesenchymal progenitor cells. As used herein, the phrase "STRO-1+ pluripotent cells" should be taken to mean STRO-1+ and / or TNAP+ progenitor cells that are capable of forming pluripotent cell colonies.

[0044] STRO-1+ pluripotent cells are cells found in bone marrow, blood, dental pulp cells, adipose tissue, skin, spleen, pancreas, brain, kidney, liver, heart, retina, brain, hair follicles, intestine, lung, lymph nodes, thymus, bone, ligaments, tendons, skeletal muscle, dermis, and periosteum, and can differentiate into germ lineages such as mesoderm and / or endoderm and / or ectoderm. Thus, STRO-1+ pluripotent cells can differentiate into numerous cell types, including, but not limited to, adipose tissue, bone tissue, cartilage tissue, elastic tissue, muscle tissue, and fibrous connective tissue. The specific lineage commitment and differentiation pathways these cells enter depend on various influences from mechanical influences and / or endogenous bioactive factors, such as growth factors, cytokines, and / or local microenvironmental conditions established by the host tissue.

[0045] Mesenchymal progenitor or stem cells can be isolated from host tissue and enriched by selecting for STRO-1+ cells. For example, bone marrow aspirate from a subject can be further treated with antibodies against STRO-1 or TNAP to allow for the selection of mesenchymal progenitor or stem cells. In one example, mesenchymal progenitor or stem cells can be enriched using STRO-1 antibodies as described in (Simmons and Torok-Storb, 1991).

[0046] The terms "enriched," "enrichment," or variations thereof, are used herein to describe a population of cells in which the population of cells of one particular cell type or the population of multiple particular cell types is increased when compared to an untreated population of cells (e.g., cells in their natural environment). In one example, a population enriched for STRO-1+ cells contains at least about 0.1%, or 0.5%, or 1%, or 2%, or 5%, or 10%, or 15%, or 20%, or 25%, or 30%, or 50%, or 75% STRO-1+ cells. In this regard, the term "population of cells enriched for STRO-1+ cells" provides explicit support for the term "a population of cells comprising X% STRO-1+ cells," where X% will be interpreted as a percentage as recited herein. STRO-1+ cells, in some examples, can form clonal colonies, e.g., CFU-F (fibroblasts), or a subset of them (e.g., 50%, 60%, 70%, 90%, or 95%) can have this activity.

[0047] In one example, a population of cells is enriched from a cell preparation that contains STRO-1+ cells in a selectable format. In this regard, the term "selectable format" will be understood to mean that the cells express a marker (e.g., a cell surface marker) that allows for the selection of STRO-1+ cells. The marker can be, but need not be, STRO-1. For example, cells (e.g., mesenchymal progenitor cells) that express STRO-2 and / or STRO-3 (TNAP) and / or STRO-4 and / or VCAM-1 and / or CD146 and / or 3G5 also express STRO-1 (and STRO-1 bright (It can be that the cells are STRO-1+. Thus, the designation that cells are STRO-1+ does not mean that the cells are selected due to STRO-1 expression. In one example, the cells are selected based on at least STRO-3 expression, e.g., they are STRO-3+ (TNAP+).

[0048] Reference to the selection of cells or populations thereof does not necessarily require selection from a specific tissue source. As described herein, STRO-1+ cells can be selected or isolated or enriched from a wide variety of sources. In some examples, these terms provide support for selection from any tissue containing STRO-1+ cells (e.g., mesenchymal progenitor cells) or vascularized tissue or tissue containing pericytes (e.g., STRO-1+ pericytes), or any one or more of the tissues listed herein.

[0049] In one example, mesenchymal progenitor or stem cells used in the present disclosure express one or more markers, individually or collectively, selected from the group consisting of TNAP+, VCAM-1+, THY-1+, STRO-2+, STRO-4+ (HSP-90β), CD45+, CD146+, 3G5+, or any combination thereof.

[0050] Use of the term "individually" means that the present disclosure encompasses the listed markers or groups of markers separately, and that notwithstanding that individual markers or groups of markers may not be individually described herein, the appended claims may define such markers or groups of markers separately and separably from one another.

[0051] Use of the term "collectively" means that the disclosure encompasses any number or combination of the listed markers or groups of markers, and that although such number or combination of markers or groups of markers may not be specifically set forth herein, the appended claims may define such combinations or sub-combinations separately and separably from any other combinations of markers or groups of markers.

[0052] In one example, STRO-1+ cells are bright (Synonym STRO-1 bri ) In another example, STRO-1 briSTRO-1 cells dim or STRO-1 intermediate In another example, STRO-1 bri The cells may further be one or more of TNAP+, VCAM-1+, THY-1+, STRO-2+, STRO-4+ (HSP-90β), and / or CD146+. For example, the cells may be selected for and / or shown to express one or more of the above-mentioned markers. In this regard, cells shown to express a marker need not be specifically tested; rather, previously enriched or isolated cells can be tested, and it can be reasonably assumed that subsequently used, isolated, or enriched cells will also express the same markers.

[0053] In one example, the mesenchymal precursor cells are perivascular mesenchymal precursor cells as defined in WO2004 / 85630, characterized by the presence of the perivascular marker 3G5.

[0054] Cells referred to as "positive" for a given marker may express either low (lo or dim) or high (bright, bri) levels of that marker, depending on the degree to which the marker is present on the cell surface; the terms refer to the intensity of the fluorescence or other marker used in the cell sorting process. The distinction between lo (or dim or dull) and bri will be understood in the context of the marker used in the particular cell population being sorted. Cells referred to as "negative" for a given marker do not necessarily mean that the marker is completely absent from that cell. This term means that the marker is expressed at a relatively very low level by the cell and, when detectably labeled or not detected above background levels, e.g., the level detected using an isotype control antibody, it produces a very low signal.

[0055] As used herein, the terms "bright" or "bri" refer to a marker on the cell surface that, when detectably labeled, produces a relatively large signal. Without wishing to be bound by theory, it is proposed that "bright" cells express more of the target marker protein (e.g., an antibody recognized by STRO-1) than other cells in the sample. For example, STRO-1 bri Cells were labeled with FITC-conjugated STRO-1 antibody as measured by fluorescence-activated cell sorting (FACS) analysis, resulting in low-brightness cells (STRO-1 dull / dim ) produce a greater fluorescent signal than STRO-1. In one example, the "bright" cells comprise at least about 0.1% of the most brightly labeled bone marrow mononuclear cells contained in the starting sample. In other examples, the "bright" cells comprise at least about 0.5%, at least about 1%, at least about 1.5%, or at least about 2% of the most brightly labeled bone marrow mononuclear cells contained in the starting sample. In one example, STRO-1 bright The cells have a 2 log (magnitude) higher expression of STRO-1 surface expression compared to "background", i.e., cells that are STRO-1-. dim and / or STRO-1 intermediate The cells have high expression, less than 2 logs (magnitude) of STRO-1 surface expression, usually about 1 log or below "background."

[0056] As used herein, the term "TNAP" is intended to encompass all isoforms of tissue non-specific alkaline phosphatase. For example, the term encompasses liver isoform (LAP), bone isoform (BAP), and kidney isoform (KAP). In one example, TNAP is BAP. In one example, TNAP as used herein refers to a molecule capable of binding to STRO-3 antibody produced by the hybridoma cell line deposited with ATCC under the provisions of the Budapest Treaty on December 19, 2005 under deposit accession number PTA-7282.

[0057] Additionally, in one example, STRO-1+ cells are capable of giving rise to clonogenic CFU-F.

[0058] In one example, a significant proportion of STRO-1+ pluripotent cells are capable of differentiation into at least two different germ cell lineages. Non-limiting examples of lineages to which pluripotent cells can be committed include bone progenitor cells; multipotent stem cell progenitors to bile duct epithelial cells and stem cells; neural restricted cells that can give rise to glial cell precursors that progress to oligodendrocytes and astrocytes; neural precursors that progress to neurons; precursors to cardiac muscle and cardiomyocytes; and glucose-responsive insulin-secreting pancreatic beta cell lines. Other lineages include, but are not limited to, odontoblasts, dentin-producing cells, and chondrocytes, as well as the following progenitor cells: retinal pigment epithelial cells, fibroblasts, skin cells such as keratinocytes, dendritic cells, hair follicle cells, renal duct epithelial cells, smooth muscle cells and skeletal muscle cells, testicular progenitor cells, vascular endothelial cells, tendon cells, ligament cells, cartilage tissue cells, adipocytes, fibroblasts, bone marrow stromal cells, cardiac muscle cells, smooth muscle cells, skeletal muscle cells, pericytes, vascular cells, glial cells, neurons, astrocytes, and oligodendrocytes.

[0059] In embodiments of the present disclosure, the mesenchymal progenitor or stem cells described herein are MSCs. The MSCs may be a homogenous composition or a mixed cell population enriched for MSCs. Homogeneous MSC cell compositions may be obtained by culturing adherent bone marrow or periosteal cells, and MSCs may be identified by specific cell surface markers identified with unique monoclonal antibodies. Methods for obtaining MSC-enriched cell populations are described, for example, in U.S. Patent No. 5,486,359. Alternative sources for MSCs include, but are not limited to, blood, skin, umbilical cord blood, muscle, fat, bone, and perichondrium.

[0060] In another example, the mesenchymal progenitor or stem cell is a CD29+, CD54+, CD73+, CD90+, CD102+, CD105+, CD106+, CD166+, MHC1+ MSC (e.g., Remestemcell-L).

[0061] The isolated or enriched mesenchymal progenitor or stem cells can be expanded in vitro by culture. The isolated or enriched mesenchymal progenitor or stem cells can be cryopreserved, thawed, and subsequently expanded in vitro by culture.

[0062] In one example, isolated or enriched mesenchymal progenitor or stem cells are cultured in a medium (serum-free or serum-supplemented), such as alpha minimal essential medium (αMEM) supplemented with 5% fetal bovine serum (FBS) and glutamine, at a density of 50,000 viable cells / cm. 2 Cells are seeded in a medium container and allowed to adhere overnight at 37°C and 20% O. The medium is then replaced and / or changed as needed, and the cells are cultured for an additional 68-72 hours at 37°C and 5% O.

[0063] As will be appreciated by those skilled in the art, cultured mesenchymal progenitor or stem cells are phenotypically distinct from in vivo cells. For example, in one embodiment, they express one or more of the following markers: CD44, NG2, DC146, and CD140b. Cultured mesenchymal progenitor or stem cells are also biologically distinct from in vivo cells, having a higher proliferation rate compared to in vivo, primarily non-cycling (quiescent) cells.

[0064] The mesenchymal precursor or stem cells may also be cryopreserved prior to administration to a subject.

[0065] Ang1 and / or VEGF expression The mesenchymal precursor or stem cells of the present disclosure can be genetically modified or genetically unmodified and express high levels of Ang1. For example, the mesenchymal precursor or stem cells can be 10 6In other examples, the cells can express an amount of Ang1 of at least 0.1 μg per cell. 6 At least 0.2 μg per cell, 10 6 0.3 μg per cell, 10 6 0.4 μg per cell, 10 6 0.5 μg per cell, 10 6 0.6 μg per cell, 10 6 0.7 μg per cell, 10 6 0.8 μg per cell, 10 6 0.9 μg per cell, 10 6 1 μg per cell, 10 6 1.1 μg per cell, 10 6 1.2 μg per cell, 10 6 1.3 μg per cell, 10 6 1.4 μg per cell, 10 6 Ang1 may be expressed in amounts of 1.5 μg per cell.

[0066] In another example, a population of mesenchymal progenitor or stem cells is 6 In other examples, the cells express VEGF in an amount of less than about 0.05 μg per cell. 6 Approximately 0.05 μg per cell, 10 6 0.04 μg per cell, 10 6 0.03 μg per cell, 10 6 0.02 μg per cell, 10 6 0.01 μg per cell, 10 6 0.009 μg per cell, 10 6 0.008 μg per cell, 10 6 0.007 μg per cell, 10 6 0.006 μg per cell, 10 6 0.005 μg per cell, 10 6 0.004 μg per cell, 10 6 0.003 μg per cell, 10 6 0.002 μg per cell, 10 6 They express less than 0.001 μg of VEGF per cell.

[0067] In another example, the mesenchymal progenitor or stem cells express Ang1:VEGF in a ratio of at least about 2: 1. In other examples, the cells express Ang1:VEGF in a ratio of at least about 10:1, 15:1, 20:1, 21:1, 22:1, 23:1, 24:1, 25:1, 26:1, 27:1, 28:1, 29:1, 30:1, 31:1, 32:1.

[0068] In some instances, the mesenchymal precursor or stem cells are genetically unmodified and express the above-described levels of Ang-1 or VEGF, or the above-described ratios of Ang-1:VEGF. As used herein, the term "genetically unmodified" refers to cells that have not been modified by transfection with a nucleic acid. For the avoidance of doubt, in the context of the present disclosure, mesenchymal precursor or stem cells transfected with a nucleic acid encoding Ang1 will be considered genetically modified.

[0069] The amount of cellular Ang1 and / or VEGF expressed in culture or present in a composition of mesenchymal progenitor or stem cells may be measured by a variety of methods known to those skilled in the art, including, but not limited to, Western blot, enzyme-linked immunosorbent assay (ELISA), fluorescence-linked immunosorbent assay (FLISA), competitive assay, radioimmunoassay, lateral flow immunoassay, flow-through immunoassay, electrochemiluminescence assay, nephelometry-based assay, turbidity-based assay, fluorescence-activated cell sorting (FACS)-based assay for detection of Ang-1 or VEGF in the medium used to culture mesenchymal progenitor or stem cells, and surface plasmon resonance (SPR or Biacore).

[0070] In one example, the levels of Ang1 and / or VEGF expressed in culture or present in a composition of mesenchymal progenitor cells or stem cells are measured in an ELISA assay. For example, cell lysates from a culture of mesenchymal progenitor cells or stem cells are added to the wells of an ELISA plate. The wells may be coated with a primary antibody, either monoclonal or polyclonal, against Ang1 or VEGF. The wells are washed and then contacted with a secondary antibody, either monoclonal or polyclonal, against the primary antibody. The secondary antibody is conjugated to a suitable enzyme, such as horseradish peroxidase. After a suitable incubation period, the wells are washed and then contacted with a suitable substrate for the enzyme conjugated to the secondary antibody, such as one or more chromogens. Chromogens that can be used include, but are not limited to, hydrogen peroxide and tetramethylbenzidine. After the substrate(s) are added, the wells are incubated for a suitable period of time. After the incubation is complete, a "stop" solution is added to the wells to stop the reaction with the enzyme's substrate(s). The optical density (OD) of the sample is then measured. The optical density of the sample to determine the amount of Ang1 or VEGF expressed by the culture of mesenchymal progenitor or stem cells being tested is correlated with the optical density of a sample containing a known amount of Ang1 or VEGF. Methods for determining the Ang1:VEGF expression ratio will also be apparent to those skilled in the art. For example, after quantifying the levels of Ang1 and VEGF, the ratio based on the quantified levels of Ang1 and VEGF can be expressed as (Ang1 level / VEGF level) = Ang1:VEGF ratio.

[0071] Methods for treating advanced heart failure Heart failure occurs when the heart cannot pump sufficiently to maintain blood flow to meet the body's needs. One cause of heart failure is systolic dysfunction after a myocardial infarction (MI). MI occurs when blood no longer flows properly to a portion of the heart. The lack of blood supply leads to a localized area of ​​myocardial necrosis called an infarction or infarction. The infarcted heart is unable to pump sufficiently to maintain blood volume to meet the body's needs, resulting in multiple pathophysiological responses, ultimately heart failure. After an MI, a series of compensatory mechanisms are initiated that help buffer the decline in cardiac output and maintain sufficient blood pressure to perfuse vital organs. As a result, patients with heart failure may not progress for a long time. However, the compensatory mechanisms eventually fail to compensate for the damaged heart, resulting in a gradual decline in cardiac output, referred to as "advanced heart failure." In the context of this disclosure, the terms chronic heart failure, congestive heart failure, congestive heart failure, systolic dysfunction, and advanced heart failure can be used interchangeably with "advanced heart failure."

[0072] The methods of the present disclosure relate to treating the gradual decline in cardiac output characteristic of progressive heart failure. Accordingly, in the context of this disclosure, "treat" and "treatment" refer to both therapeutic treatment and preventative or prophylactic measures.

[0073] In some instances, treatment reduces the likelihood or risk of heart failure-associated adverse cardiac events (HF-MACE), defined as cardiac-related death or resuscitated cardiac death, or non-fatal decompensated heart failure events. In some instances, the likelihood or risk of HF-MACE is reduced for at least 6 months, at least 12 months, at least 24 months, or at least 36 months. In some instances, treatment reduces the likelihood or risk of all-cause mortality.

[0074] Myocardial infarction subjects The term "myocardial infarction (MI) subject" is used to define a subject who has had a myocardial infarction. The methods of the present disclosure can be used to treat progressive heart failure in a specific population of MI subjects. Subjects in need of treatment include subjects who already have progressive heart failure and subjects in whom progressive heart failure should be prevented, delayed, or stopped.

[0075] MI subjects treated with the disclosed method have proximal left anterior descending (LAD) artery disease. As understood by those skilled in the art, the LAD artery runs in the anterior interventricular groove at the front of the heart, separating the right and left ventricles. The diagonal (Dx) branch emerges from the LAD and runs across the anterior wall to the outer surface, or lateral portion. Thus, the Dx supplies blood to the anterior lateral portion of the left ventricle. A subject may have one or several Dx branches. The first Dx branch serves as the boundary between the proximal and middle portions of the LAD. Thus, the portion of the artery before the origin of the Dx is known as the "proximal LAD," while the segment adjacent to the first major aspect of the branch. The distal segment of the LAD is the terminal third of the artery.

[0076] In the context of this disclosure, the term "arterial lesion" includes an occlusive lesion that occludes the LAD of the heart, or an arterial lesion that previously occluded the LAD that has been treated, for example, by percutaneous coronary intervention (PCI), also known as angioplasty.

[0077] In some instances, a subject treated with the methods of the present disclosure underwent PCI within about 1 hour of ischemic symptoms. In other instances, the subject underwent PCI within about 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 hours of ischemic symptoms. In some instances, the subject underwent PCI within about 12 hours of ischemic symptoms. In other instances, the subject underwent PCI within about 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or more hours. Subjects treated with thrombolytic therapy who have recurrent chest pain and / or ECG changes may not transition to PCI until at least 24 hours after the onset of ischemic symptoms. Thus, in other examples, the subject underwent PCI within about 24 hours, about 25 hours, about 26 hours, about 27 hours, about 28 hours, about 29 hours, about 30 hours, about 35 hours, about 40 hours, or about 48 hours of the ischemic episode.

[0078] A subject with MI may have elevated left ventricular end-systolic volume (LVESV). In some instances, a subject with MI treated using the methods of the present disclosure has an elevated LVESV of more than 70 mL. In some instances, the subject has an elevated LVESV of more than 80 mL, more than 90 mL, more than 100 mL, more than 110 mL, or more than 120 mL. In other instances, the subject has an elevated LVESV of more than 80 mL / m 2 More than 90 mL / m 2 More than 100 mL / m 2 exceeding 110 mL / m 2 or more than 120 mL / m 2 have an elevated LVESV of more than

[0079] MI can cause persistent left ventricular dysfunction. Thus, in another example, a subject with MI has proximal LAD artery disease and persistent left ventricular dysfunction. Left ventricular dysfunction is characterized by a decrease in myocardial contractility. When the myocardial contractility in the left ventricle decreases, a decrease in left ventricular ejection fraction (LVEF) occurs. Thus, LVEF provides a means of determining left ventricular dysfunction.

[0080] LVEF and LVESV can be measured by a number of methods known in the art, such as echocardiogram, single photon emission computed tomography (SPECT), or cardiac magnetic resonance imaging (cMRI).

[0081] In some examples, subjects with an LVEF of less than about 60% have left ventricular dysfunction. In other examples, subjects with an LVEF of less than about 55%, 54%, 53%, 52%, 51%, 50%, 49%, 48%, 47%, or 46% have left ventricular dysfunction. In another example, subjects with an LVEF of less than about 45% have left ventricular dysfunction. In another example, subjects with an LVEF of less than about 44%, 43%, 42%, or 41% have left ventricular dysfunction. In another example, subjects with an LVEF of less than about 40% have left ventricular dysfunction. In another example, subjects with an LVEF of less than about 39%, 38%, 37%, 36%, 35%, 34%, 33%, 32%, 31%, or 30% have left ventricular dysfunction.

[0082] In the context of the present disclosure, the term "persistent left ventricular dysfunction" is used to define left ventricular dysfunction that persists for a certain period of time or series of measurements. For example, "persistent left ventricular dysfunction" can include left ventricular dysfunction that persists for about 1 to about 14 days or longer after MI. For example, persistent left ventricular dysfunction can include left ventricular dysfunction that persists for about 1 to about 10 days, about 1 to about 9 days, about 2 to about 8 days, or about 2 to about 7 days after MI. In another example, "persistent left ventricular dysfunction" can include left ventricular dysfunction that persists for about 1 to 10 or more measurements.

[0083] The size or amount of myocardial necrosis after an MI is clinically referred to as infarct size. The methods of the present disclosure relate to treating subjects with an MI who have a large infarct size. For example, subjects treated using the methods of the present disclosure can have an infarct size of greater than about 10-35% of the left ventricle. In other examples, subjects have an infarct size of greater than about 11-34%, about 12-33%, about 13-32%, about 14-31%, about 15-30%, about 16-29%, or about 17-28% of the left ventricle. In other examples, subjects have an infarct size of greater than about 18.5% of the left ventricle. In other examples, subjects have an infarct size of greater than about 19-27%, about 20-26%, about 21-25%, about 22-24%, or about 23% of the left ventricle.

[0084] Infarct size can be measured by a number of methods known in the art, including the use of serum markers such as creatine kinase (CK), CK-MB, troponin I, and brain natriuretic peptide troponin.

[0085] In some instances, subjects treated with the methods of the present disclosure have troponin levels at least about 2 times the upper limit of normal (ULM). In other instances, subjects have troponin levels at least about 3 times, 4 times, 5 times, or 6 times the ULM.

[0086] In some examples, a subject treated with the methods of the present disclosure has creatine kinase-MB levels that are at least about 2-fold higher than ULM. In other examples, the subject has creatine kinase-MB levels that are at least about 3-fold, 4-fold, 5-fold, or 6-fold higher than ULM.

[0087] Other examples of measuring infarct size include sestamibi single-photon emission computed tomography (SPECT), myocardial perfusion imaging, and magnetic resonance imaging. In one example, infarct size is measured using cMRI. Several cMRI techniques can be used to diagnose infarct size. One of the most accurate and well-validated techniques is delayed-enhancement cardiac magnetic resonance imaging (DE-CMR). Thus, in one example, cMRI includes DE-CMR.

[0088] When appropriate settings for DE-CMR are used, normal myocardium appears black or empty, while nonviable areas appear bright or hyperenhanced. Thus, in some instances, infarct size can be measured by visual assessment of bright, hyperenhanced areas. Other methods for determining infarct size are known in the art (Sievers et al. (2007), Circulation, 115, 236-244; Kim et al. (2000), N Engl J Med, 343, 1445-1453). Briefly, hyperenhancement is scored in a 17-segment model using a 5-point scale for each segment (0 = no hyperenhancement, 1 = 1%-25%, 2 = 26%-50%, 3 = 51%-75%, 4 = 76%-100%). Dark regions completely contained within the highly enhancing myocardium are interpreted as regions of microvascular damage (no-reflow) and included as part of the infarct. Infarct size as percent LV myocardium is calculated by summing the regional scores, each weighted by the midpoint of the hyperenhancement range (i.e., 1=13%, 2=38%, 3=63%, 4=88%), and dividing by 17. In another example, infarct size can be quantified by planimetry of the highly enhancing area on a stack of short-axis images.

[0089] In some instances, infarct size is measured between about 1 and 40 days after MI. In other instances, infarct size is measured between about 1 and 40 days, between about 2 and 35 days, between about 3 and 30 days, between about 4 and 25 days, between about 5 and 20 days, or between about 6 and 15 days after MI. For example, infarct size can be measured about 30 days after MI.

[0090] In the context of the present disclosure, "infarct size" refers to left ventricular infarct size. In other words, left ventricular infarct size refers to the volume of the left ventricle that is infarcted.

[0091] The disclosed methods can be used to treat progressive heart failure in subjects with MI at various stages or classifications of heart failure. For example, the subject can have stage A, B, C, or D heart failure. In some examples, the subject has stage B or C heart failure. In these examples, the staging of heart failure is based on the staging criteria of the American College of Cardiology (ACC) and the American Heart Association (AHA).

[0092] In another example, the subject can have Class I, II, III, or IV heart failure. In some examples, the subject has Class II or III heart failure. In these examples, the heart failure classification is based on the New York Heart Association (NYHA) classification scale.

[0093] cell composition In practicing the methods of the present disclosure, the mesenchymal progenitor or stem cells can be administered in the form of a composition, hi one example, such a composition includes a pharmaceutically acceptable carrier and / or excipient.

[0094] The terms "carrier" and "excipient" refer to compositions of substances conventionally used in the art to facilitate the storage, administration, and / or biological activity of active compounds (see, e.g., Remington's Pharmaceutical Sciences, 16th Ed., Mac Publishing Company (1980)). A carrier may also reduce any undesirable side effects of the active compound. Suitable carriers are, for example, stable and, for example, incapable of reacting with other ingredients in the carrier. In one example, a carrier does not produce local or systemic adverse reactions in the recipient at the dosages and concentrations used in treatment.

[0095] Suitable carriers for the present disclosure include those conventionally used, for example, water, saline, aqueous dextrose, lactose, Ringer's solution, buffer, hyaluronan, and glycols are exemplary liquid carriers, particularly for solutions (when isotonic). Suitable pharmaceutical carriers and excipients include starch, cellulose, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, magnesium stearate, sodium stearate, glycerol monostearate, sodium chloride, glycerol, propylene glycol, water, ethanol, and the like.

[0096] In another example, the carrier is, for example, a media composition that the cells grow or thrive in. Such a media composition does not cause any adverse effects to the subject to which it is administered.

[0097] Exemplary carriers and excipients do not adversely affect cell viability and / or the ability of the cells to reduce, prevent, or delay progressive heart failure.

[0098] In one example, the carrier or excipient provides buffering activity and / or biological activity to maintain the cells and soluble factors at a suitable pH, thereby exerting their biological activity; for example, the carrier or excipient is phosphate buffered saline (PBS). PBS represents an attractive carrier or excipient because it interacts minimally with the cells and factors, allowing for rapid release of the cells and factors. In this case, the compositions of the present disclosure may be formulated as a liquid for application, for example, by injection, to the bloodstream or directly to tissues or areas surrounding or adjacent to the tissues.

[0099] These stem cells and / or progeny cells can also be incorporated or embedded into scaffolds that degrade into products that are compatible with and not harmful to the recipient. These scaffolds provide support and protection to the cells to be transplanted into the recipient subject. Examples of such scaffolds are natural and / or synthetic biodegradable scaffolds.

[0100] A variety of different scaffolds may be successfully used in the practice of the present disclosure. Exemplary scaffolds include, but are not limited to, biodegradable scaffolds. Natural biodegradable scaffolds include collagen, fibronectin, and laminin scaffolds. Synthetic materials suitable for cell transplantation scaffolds should be able to support a wide range of cell growth and function. Such scaffolds may also be resorbable. Suitable scaffolds include polyglycolic acid scaffolds (e.g., as described by Vacanti, et al. J. Ped. Surg. 23:3-9 1988; Cima, et al. Biotechnol. Bioeng. 38:145 1991; Vacanti, et al. Plast. Reconstr. Surg. 88:753-9 1991); or synthetic polymers such as polyanhydrides, polyorthoesters, and polylactic acid.

[0101] In another example, cells may be administered in a gel scaffold (eg, Gelfoam from Upjohn Company).

[0102] The cell compositions described herein may be administered alone or as a mixture with other cells. Different types of cells may be mixed with the compositions of the present disclosure immediately or immediately prior to administration, or they may be co-cultured together for a period of time prior to administration.

[0103] In one example, the composition comprises an effective amount or a therapeutically or prophylactically effective amount of cells. For example, the composition comprises about 1 x 10 5 cells ~ approx. 1×10 9 cells or approximately 1.25 x 10 3 Cells ~ approx. 1.25×10 7 The exact amount of cells to be administered will depend on a variety of factors, including the age, weight, and sex of the subject, as well as the extent and severity of the disorder being treated.

[0104] An exemplary dosage is at least about 1.2 x 10 8 ~Approx. 8×10 10cells, e.g., about 1.3 x 10 8 ~Approx. 8×10 9 cells, approximately 1.4 x 10 8 ~Approx. 8×10 8 cells, approximately 1.5 x 10 8 ~Approx. 7.2×10 8 cells, approximately 1.6 x 10 8 ~Approx. 6.4×10 8 cells, approximately 1.7 x 10 8 ~Approx. 5.6×10 8 cells, approximately 1.8 x 10 8 ~Approx. 4.8×10 8 cells, approximately 1.9 x 10 8 ~Approx. 4.0×10 8 cells, approximately 2.0 x 10 8 ~Approx. 3.2×10 8 cells, approximately 2.1 x 10 8 ~Approx. 2.4×10 8 For example, the dosage may be at least about 1.5 x 10 cells. 8 For example, the dosage may include at least about 2.0 x 10 cells. 8 It may contain cells.

[0105] In other words, an exemplary dosage is at least about 1.5 x 10 per kg (for an 80 kg subject). 6 In some instances, the dosage is at least about 2.5 x 10 cells per kg. 6 In another example, the dosage may be about 1.5 x 10 cells per kg. 6 ~Approx. 1×10 9 Approximately 1.6 x 10 cells per kg 6 ~Approx. 1×10 8 Approximately 1.8 x 10 cells per kg 6 ~Approx. 1×10 7 Approximately 1.9 × 10 cells per kg 6 ~Approx. 9×10 6 Approximately 2.0 × 10 cells per kg 6 ~Approx. 8×10 6 Approximately 2.1 x 10 cells per kg 6 ~Approx. 7×10 6 Approximately 2.3 x 10 cells per kg 6 ~Approx. 6×10 6Approximately 2.4 x 10 cells per kg 6 ~Approx. 5×10 6 Approximately 2.5 × 10 cells per kg 6 ~Approx. 4×10 6 Approximately 2.6 x 10 cells per kg 6 ~Approx. 3×10 6 This may include between cells.

[0106] In some examples, the mesenchymal progenitor or stem cells comprise at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 99% of the cell population of the composition.

[0107] The compositions of the present disclosure may be cryopreserved. Cryopreservation of mesenchymal progenitor or stem cells can be performed using slow cooling methods or "rapid" freezing protocols known in the art. Preferably, the cryopreservation method maintains a similar phenotype, cell surface markers, and growth rate of the cryopreserved cells compared to unfrozen cells.

[0108] The cryopreservation composition may contain a cryopreservation solution, the pH of which is usually 6.5 to 8, preferably 7.4.

[0109] Cryopreservation solutions may include, for example, sterile, non-pyrogenic, isotonic solutions such as PlasmaLyte A™. 100 mL of PlasmaLyte A™ contains 526 mg of sodium chloride, USP (NaCl); 502 mg of sodium gluconate (C6H 11NaO7); 368 mg of sodium acetate trihydrate, USP (C2H3NaO2 3H2O); 37 mg of potassium chloride, USP (KCl); and 30 mg of magnesium chloride, USP (MgCl2 6H2O). It contains no antimicrobial agents. The pH is adjusted with sodium hydroxide. The pH is 7.4 (6.5-8.0).

[0110] The cryopreservation solution may include Profreeze™. The cryopreservation solution may additionally or alternatively include a culture medium, such as αMEM.

[0111] To facilitate freezing, cryoprotectants, such as dimethyl sulfoxide (DMSO), are typically added to cryopreservation solutions. Ideally, cryoprotectants should be nontoxic to cells and the host, nonantigenic, and chemically inert, provide high post-thaw survival rates, and allow for transplantation without washing. However, the most commonly used cryoprotectant, DMSO, exhibits some cytotoxicity. Hydroxyethyl starch (HES) may be used as a substitute or in combination with DMSO to reduce the cytotoxicity of cryopreservation solutions.

[0112] The cryopreservation solution may include one or more of DMSO, hydroxyethyl starch, human serum components, and other protein bulking agents. In one example, the cryopreserved solution includes about 5% human serum albumin (HSA) and about 10% DMSO. The cryopreservation solution may further include one or more of methylcellulose, pyrrolidone (PVP), and trehalose.

[0113] In one embodiment, cells are suspended in 42.5% Profreeze™ / 50% αMEM / 7.5% DMSO and cooled in a controlled rate freezer.

[0114] The cryopreserved composition may be thawed and administered directly to a subject, or added to another solution containing, for example, HA. Alternatively, the cryopreserved composition may be thawed and the mesenchymal progenitor or stem cells resuspended in an alternative carrier prior to administration.

[0115] In some instances, the cell compositions described herein may be administered between about 1 and about 10 days post-MI. In other instances, the cell compositions described herein may be administered between about 1 and 9 days, between about 1 and 8 days, between about 2 and 7 days, between about 2 and 6 days, or between about 3 and 5 days post-MI. For example, the cell compositions described herein may be administered about 5 days post-MI.

[0116] In some instances, the cell compositions described herein may be administered between about 1 and about 10 days after PCI. In other instances, the cell compositions described herein may be administered between about 1 and 9 days, between about 1 and 8 days, between about 2 and 7 days, between about 2 and 6 days, or between about 3 and 5 days after PCI. For example, the cell compositions described herein may be administered about 5 days after PCI.

[0117] In some instances, the cell compositions described herein may be administered as a single dose. In other instances, the cell compositions are administered over multiple doses, such as at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 doses.

[0118] In one example, mesenchymal precursor cells or stem cells can be expanded before administration. Various methods for culturing mesenchymal precursor cells or stem cells are known in the art. In one example, mesenchymal precursor cells or cells are expanded in serum-free medium before administration.

[0119] Mesenchymal progenitor cells or stem cells may be administered systemically, for example, intravenously, intraarterially, or intraperitoneally. Mesenchymal progenitor cells or stem cells may also be administered intranasally, intramuscularly, or intracardially. In some instances, mesenchymal progenitor cells or stem cells are administered directly into the myocardium. For example, mesenchymal progenitor cells or stem cells can be administered directly into the myocardium of the left ventricle. In some instances, mesenchymal progenitor cells or stem cells are administered via an endocardial catheter, such as a J&J Myostar™ injection catheter.

[0120] In some instances, mesenchymal progenitor cells or stem cells are administered to viable myocardium. In some instances, mesenchymal progenitor cells or stem cells are administered to hibernating myocardium. One skilled in the art would be able to identify viable and / or hibernating myocardium using methods known in the art. For example, a mapping catheter system, such as the NOGASTAR™ mapping catheter system, can be used to identify viable and / or hibernating myocardium.

[0121] In another example, mesenchymal progenitor cells or stem cells are administered by intracoronary injection.For example, mesenchymal progenitor cells or stem cells can be administered to the left anterior descending artery (LAD).In one example, mesenchymal progenitor cells or stem cells are administered to the LAD artery immediately after PCI LAD revascularization.

[0122] In some instances, the cells are contained within a chamber that does not allow the cells to exit the subject's circulation but allows factors secreted by the cells to enter the circulation. In this manner, a soluble factor may be administered to a subject by allowing the cells to secrete the factor into the subject's circulation. Such a chamber may equally be implanted at a site in a subject, for example, in or near the heart, to increase the local level of the soluble factor.

[0123] It will be appreciated by those skilled in the art that numerous variations and / or modifications may be made to the present invention as illustrated in the specific embodiments without departing from the spirit or scope of the invention as broadly described. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive.

[0124] All publications discussed and / or referenced herein are incorporated herein in their entirety.

[0125] Any discussion of documents, acts, materials, devices, articles or the like which has been included in the present specification is solely for the purpose of providing a context for the present invention and is not to be construed as an admission that any or all of such matter forms part of the prior art base or was common general knowledge in the art relevant to the present invention as existing before the priority date of each claim of this application.

[0126] This application claims priority from AU 2014905240, filed December 23, 2014, the disclosure of which is incorporated herein by reference. [Example]

[0127] Example 1 MSC in clinical phase 2a / 2b IV AMI A randomized, placebo-controlled phase 2a / 2b clinical trial has been initiated to evaluate a single intravenous (IV) dose of 200 million mesenchymal stem cells (MSCs) administered over 2–7 days in patients with ST-elevation MI (STEMI) or non-ST-elevation myocardial infarction (NSTEMI) and reduced cardiac ejection fraction.

[0128] A phase IIa / IIb, multicenter, randomized, double-blind, placebo-controlled study was designed to evaluate the safety and efficacy of intravenous infusion of Remestemcel-L (ex vivo cultured adult human mesenchymal stem cells) after acute myocardial infarction.

[0129] The study was a phase II, multicenter, randomized, double-blind, placebo-controlled study of 220 subjects who had recently experienced an acute myocardial infarction (MI). Patients with acute MI were eligible for the study: 1) positive biomarkers (troponin or CK-MB >4x ULN); 2) regional wall motion abnormalities; 3) Patients were included who had any acute coronary syndrome resulting in a decline in global left ventricular systolic function of 45% or less and 20% or more as determined by screening cardiac imaging performed within approximately 24 hours after the initial onset of the acute event.

[0130] Subjects were evaluated for safety and efficacy until death, discontinuation, or 60 months after investigational drug (IA) infusion, whichever occurred first. The treatment window was 2 to 7 days after the first episode of acute MI.

[0131] Subjects were randomly assigned to placebo or treatment groups (200 × 10 Remestemcel-L per injection). 6 Approximately equal numbers of subjects in each cohort were assigned to each of the two groups (1:1 randomization).

[0132] Full study data over 24 months All intravenous mesenchymal stem cell (MSC) treatments were well tolerated, and studies demonstrated that the use of IV MSCs during acute myocardial infarction was safe. There were no clear differences between remestemcel-L and placebo in total adverse events (90.9% vs. 90.9%) and serious adverse events (32.7% vs. 33.6%) in subjects with acute MI. There were five deaths during the study: two in the remestemcel-L group and three in the placebo group. Only one subject's death was considered possibly related to study treatment; the subject received placebo.

[0133] In the original study design, the primary inclusion criterion for study participation was an LVEF of less than 45% as determined by cardiac imaging approximately 24 hours after the acute event; more than 70% of eligible subjects at screening were found to have an LVEF of greater than 45% by cMR immediately prior to intravenous administration of the study product, administered at an average time of 5.4 days after the MI. This is most likely due to the natural history of myocardial recovery after the MI and angioplasty procedure. Therefore, fewer than 30% of enrolled patients actually met the study design criterion of persistent left ventricular (LV) dysfunction at the time of remestemcel-L infusion; therefore, most patients in this study actually had normal LV function at the time of treatment.

[0134] Sixty-six patients had left anterior descending artery (LAD) disease with low ejection fraction at screening. For the primary efficacy variable of cMR-derived change in left ventricular end-systolic volume at 3 months, subjects in the Remestemcel-L group demonstrated a numerically non-statistically significant change (+3.31 mL vs. -0.35 mL, p=0.17). Similar non-significant numerical changes were also evident in ESV and mean infarct size and ejection fraction at 6 months.

[0135] Next, we evaluated the effect of remestemcel-L therapy in a subset of patients with persistent LV dysfunction and proximal LAD lesions, as measured by cMR immediately before infusion.

[0136] Post-hoc efficacy analysis The central rationale for this study was that Remestemcel-L would be effective in post-MI patients with persistent LV dysfunction. However, in over 70% of the overall patient population, ejection fraction normalized between inclusion of the study inclusion 24 hours after MI and treatment on days 2 to 7 (mean treatment application 5.4 days). This may reflect early recovery of stunned myocardium within days after PCI reperfusion, resulting in improved LV systolic function before treatment application. Discrepancies in LVEF data between study entry and treatment application significantly negated the original trial's ability to adequately test the study's underlying hypotheses.

[0137] Therefore, a post-hoc analysis was designed to further explore the potential effect of remestemcel-L in the subset of patients who would be expected to have the most extensive disease after MI, specifically, subjects with a proximal LAD culprit lesion and a cMR LVEF of 45% or less at the time of treatment administration.

[0138] In patients with the highest risk of advanced heart failure, a single dose (200 × 10 6 A post-hoc analysis was designed to answer whether intravenously administered MSCs (MSCs) were more effective than placebo in reversing left ventricular heart failure and preventing progressive adverse LV remodeling 6 months after the index AMI. Patients at highest risk were selected for study participation as follows: First anterior acute myocardial infarction caused by a proximal LAD culprit lesion; The culprit coronary artery lesion was successfully treated with a PCI procedure within 12 hours of the onset of ischemic symptoms. Persistent LV systolic dysfunction (i.e., post-PCI LVEF <45% due to cMR) is present 2–7 days after the index AMI.

[0139] Of the 220 patients randomized to the entire trial, a total of 25 subjects met the following criteria: MI localized to the proximal LAD, ischemic time less than 12 hours, successful percutaneous coronary intervention (PCI), baseline cMR LVEF less than 45%, and treatment with the investigational product between 2 and 7 days after the index AMI.

[0140] These 25 subjects constituted the evaluation population for the post-hoc analysis, distributed as follows: 10 subjects in the remestemcel-L group and 15 subjects in the placebo group. A post-hoc analysis conducted for the phase 2 remestemcel-L AMI trial used a responder index approach to evaluate the effect of cell therapy on adverse LV remodeling. The primary endpoint of the post-hoc study, which assessed the change from baseline in LVESV to determine whether a clinically meaningful difference existed between patients treated with remestemcel-L and placebo 6 months after AMI, was evaluated using this approach.

[0141] Of the commonly evaluated measures of LV systolic function, LVESV has previously been shown to be a strong predictor of long-term survival after recovery from acute myocardial infarction (White et al. (1987) Circulation, 76:44-51).

[0142] Indeed, it is now well recognized that LVESV is a useful surrogate efficacy endpoint for biologically and clinically meaningful changes associated with adverse LV remodeling and the associated development of MACE in patients at risk for the development / progression of heart failure related to LV systolic function. This analytical model has been shown to correlate well with MACE outcomes in phase 2 gene therapy trials (Hajjar et al. (2008) J Card Fail., 14(5):355-67; ​​Jaski et al. (2009) Card Fail., 15(3):171-81; Jessup et al. (2011) Circulation, 124:304-313).

[0143] In the current analysis of the primary and secondary endpoints, pre-specified threshold boundaries for success were used to determine treatment responders. These values, calculated as 6-month minus baseline data, were used to ensure that changes in intervals were both outside the normal range of measurement error and potentially clinically significant.

[0144] [Table 1]

[0145] Post-hoc group efficacy results At baseline, there were no significant differences between treatment groups with respect to LVESV, LVEDV, LVEF, or LV infarct volume. However, interesting trends were evident within the changes from baseline data to the 6-month follow-up visit.

[0146] LV infarct volume; At baseline, mean infarct volumes were similar and very high between the placebo and remestemcel-L groups: 40.9 ± 13.75 g (mean ± standard deviation) and 40.76 ± 17.82 g (mean ± standard deviation), respectively. Assuming a total mean myocardial mass of approximately 130 g in these patients (Stone et al. (2012) JAMA. 307:17, 1817-26), these values ​​represent a very high-risk population with an infarct size of approximately 30% at the time of therapeutic intervention. Because prospective studies have shown that an infarct size greater than 18.5% results in a 30% incidence of major adverse cardiac events (HF-MACE, defined as heart failure hospitalization or death) over 2 years, this suggests that the population of AMI patients evaluated in this post-hoc analysis was at highest risk for subsequent HF-MACE.

[0147] At the end of 6 months, subjects in the remestemcel-L group showed a reduction in LV infarct volume from a baseline value of 40.76 ± 17.82 g to 26.6 ± 12.94 g (mean ± standard deviation). This represented a change of -14.14 ± 13.94 g. Meanwhile, placebo subjects showed a smaller than nominal reduction in LV infarct volume from a baseline value of 40.9 ± 13.75 g (mean ± standard deviation) to 31.59 ± 12.70 g. This represented a change of -7.74 ± 10.84 g. The placebo-corrected difference was -6.40 g (p = 0.187).

[0148] These results indicate that remestemcel-L enhanced the natural endogenous healing process, resulting in a two-fold reduction in infarct volume over 6 months compared to placebo (assuming a mean LV mass of 130 g), from approximately 30% infarct size at baseline to approximately 20% infarct size at 6 months.

[0149] LVESV: At the end of 6 months, subjects in the remestemcel-L group demonstrated a decrease in LVESV from a baseline value of 85.0 ± 15.89 mL to 73.0 ± 24.24 mL (mean ± standard deviation). This represented a change of -12.0 ± 16.57 mL. Meanwhile, placebo subjects demonstrated an increase in LVESV from a baseline value of 90.5 ± 23.54 mL (mean ± standard deviation) to 92.8 ± 35.40 mL. This represented a change of 2.2 ± 28.53 mL. The placebo-corrected difference was -14.2 mL (p = 0.174).

[0150] [Table 2]

[0151] In particular, a change in infarct size from approximately 30% at baseline to approximately 20% at 6 months (assuming a mean LV mass of 130 g), as seen in the RemestemCell-L group, would be expected to reduce LVESV by at least 10 mL over this period (Wu et al. (2007) Stem Cells, 25:26, 48-59). Our results are consistent and confirm the concordance of our data set.

[0152] LVEDV: At the end of 6 months, subjects in the remestemcel-L group showed an increase in LVEDV from a baseline value of 142.9 ±24.01 mL to 154.4 ±37.52 mL (mean ± standard deviation), representing a change of 11.5 ±27.91 mL. Meanwhile, placebo subjects showed an increase in LVEDV from a baseline value of 151.2 ±35.98 mL (mean ± standard deviation) to 167.8 ±41.66 mL, representing a change of 16.6 ±27.30 mL. The placebo-corrected difference was -5.1 mL (p=0.618).

[0153] LVEF: At the end of 6 months, subjects in the remestemcel-L group showed an increase in LVEF from baseline values ​​of 40.6 ± 4.23% to 53.1 ± 8.71% (mean ± standard deviation), representing a change of 12.5 ± 8.88 LVEF units. Meanwhile, placebo subjects showed an increase in LVEF from baseline values ​​of 40.3 ± 3.47% (mean ± standard deviation) to 45.8 ± 9.08%, representing a change of 5.6 ± 9.48 LVEF units. The placebo-corrected difference was 6.9 LVEF units (p = 0.066) (see Figures 1 and 2).

[0154] [Table 3]

[0155] Responder analysis There was a significant trend toward statistical significance (p=0.095) for the difference between the percentage of remestemcel-L patients versus placebo patients who were treatment responders 6 months after the index AMI. Specifically, 60% of patients treated with remestemcel-L were treatment responders, compared with 27% of placebo patients. This represented a 2.2-fold increase in the responder rate for the remestemcel-L group compared with the placebo group (Figure 3).

[0156] Cohort analysis of responders vs. non-responders Although the responder rate for reduction in LVESV was 2.2 times higher in the remestemcel-L group than in the placebo group, the mean 6-month changes in parameters of LV remodeling and global LV systolic function were similar for remestemcel-L and placebo responders (reduction in LVESV, no change in LVEDV, and increase in LVEF) compared with remestemcel-L and placebo non-responders (nominal increase in LVESV, large increase in LVEDV, and minimal increase in LVEF).

[0157] In contrast, remestemcel-L treatment responder subjects demonstrated a greater reduction in LV infarct volume from baseline to Month 6 (-18.8 g) than was observed for any other subgroup (remestemcel-L non-responders = -7.1 g, placebo responders = -4.8 g, and placebo non-responders = -9.1 g). The mean change in LV infarct volume for the 17 patients included in the latter three groups was -7.6 g. This difference in LV infarct volume for remestemcel-L treatment responders contrasts markedly with the data generated for LVESV, LVEDV, and LVEF analyses; the treatment responder data were generally similar for remestemcel-L and placebo subjects.

[0158] Assuming a mean left ventricular mass of 130 g in patients with large occlusions after proximal LAD occlusion (Stone et al. (2012) JAMA., 307:17, 1817-26), this represents a change in infarct size in remestemcel-L responders from approximately 30% at baseline to approximately 17% at 6 months. This reduction in infarct volume may have a major impact on HF-MACE event rates over 2 years in this group (Wu et al. (2007) Stem Cells, 25:26, 48-59).

[0159] Therefore, remestemcel-L responders are 1) a 2.2-fold higher rate of achievement of the primary efficacy endpoint associated with LVESV (p=0.095); 2) a higher level of concordance between improved LV remodeling, improved global LV systolic function, and reduced LV infarct volume; 3) In other words, we demonstrated a unique mechanism by which reduction in infarct volume improved LVESV and adverse LV remodeling.

[0160] [Table 4]

[0161] For the overall remestemcel-L group (n=10) vs. placebo group (n=15), there were no significant differences in baseline patient demographics, time from onset of ischemic MI symptoms to time of PCI, or incidence of post-PCI TIMI perfusion flow grade 3. Compared with the placebo group, there was a trend toward shorter times from PCI to investigational product infusion and from first ischemic MI symptoms to investigational product infusion for the remestemcel-L group.

[0162] In conclusion, intravenous administration of remestemcel-L 2 to 7 days after AMI helped reduce LV infarct size, attenuate adverse LV remodeling, and improve global LV systolic function 6 months after the index event. The beneficial effects of remestemcel-L compared with placebo were evident in both the infarcted region (reduced infarct volume) and remote myocardial regions (reduced LVESV leading to increased LVEF). It is anticipated that these findings may ultimately equate to a reduced incidence of heart failure in post-AMI patients at high risk for this condition.

[0163] Example 2 Correlation between disease severity and therapeutic efficacy of MPC in LVESV Figures 4 to 9 show the results of placebo (control) or MPC (1.5 × 10 8Figure 1 shows the change in LVESV in subjects evaluated 6 months after administration of 1000kJ / kg of mesenchymal progenitor cells (MSCs). Reduction in LVESV correlated with the level of heart failure (as determined by baseline LVESV measurements). Subjects were stratified by LVESV as follows:

[0164] [Table 5]

[0165] These data demonstrate that the greater the magnitude of baseline left ventricular contractile abnormalities in subjects with chronic heart failure due to left ventricular systolic dysfunction, the more beneficial the MPC-associated cardioprotective effects observed over a 6-month follow-up period. The data further demonstrate that the progressively adverse natural history associated with progressive chronic heart failure can be beneficially altered by treatment with MPC. Without wishing to be bound by theory, the findings support the paracrine crosstalk hypothesis, in which tissue-level biochemical / physiological disturbances create a local environment that promotes MPC release of beneficial paracrine mediators. Therefore, the optimal effects achieved by administration of MPC in heart failure subjects are seen in subjects at highest risk for disease progression, i.e., those with a baseline LVESV of greater than 70 mL.

Claims

1. 1. A method for preventing progressive heart failure in a subject who has had a myocardial infarction, the method comprising administering to the subject a population of mesenchymal progenitor or stem cells and / or their progeny and / or soluble factors derived therefrom, wherein the subject has a proximal left anterior descending artery (LAD) lesion.

2. 10. The method of claim 1, wherein the subject has a LVESV of greater than 70 ml.

3. 3. The method of claim 1 or claim 2, wherein the subject has persistent left ventricular dysfunction.

4. The method of any one of claims 1 to 3, wherein the subject has a LVEF of less than about 45%.

5. The method of any one of claims 1 to 4, wherein the subject has a LVEF of less than about 40%.

6. 6. The method of any one of claims 1 to 5, wherein the mesenchymal precursor or stem cells and / or their progeny and / or soluble factors derived therefrom are administered between about 1 and 7 days after myocardial infarction.

7. 6. The method of any one of claims 1 to 5, wherein the mesenchymal precursor or stem cells and / or their progeny and / or soluble factors derived therefrom are administered between about 3 and 5 days after myocardial infarction.

8. The method of any one of claims 1 to 7, wherein the subject has creatine kinase-MB and / or troponin and / or myoglobin greater than about 4 times the upper limit of normal.

9. The method of any one of claims 1 to 8, wherein the subject has an infarct size of between about 10-25% of the left ventricle.

10. The method of any one of claims 1 to 8, wherein the subject has an infarct size greater than about 18.5% of the left ventricle.

11. The method of any one of claims 1 to 10, wherein the LVEF and / or infarct size are measured by cardiovascular magnetic resonance imaging (cMR).

12. STRO-1 ＋ 12. The method of any one of claims 1 to 11, comprising administering a population of mesenchymal precursor or stem cells enriched for cells and / or their progeny and / or soluble factors derived therefrom.

13. STRO-1 ｂｒｉｇｈｔ 13. The method of any one of claims 1 to 12, comprising administering a population of mesenchymal precursor or stem cells enriched for cells and / or their progeny and / or soluble factors derived therefrom.

14. The method of any one of claims 1 to 13, wherein the population of mesenchymal precursor or stem cells and / or their progeny and / or soluble factors derived therefrom is administered systemically.

15. 15. The method of any one of claims 1 to 14, wherein the population of mesenchymal precursor or stem cells and / or their progeny and / or soluble factors derived therefrom is administered intravenously or intranasally.

16. 16. The method of any one of claims 1 to 15, wherein said population of mesenchymal precursor or stem cells and / or their progeny and / or soluble factors derived therefrom is administered over multiple doses.

17. 1 x 10 ８ ~8 x 10 ８ The method of any one of claims 1 to 16, comprising administering cells.

18. 1.2 x 10 ８ ~4 x 10 ８ The method of any one of claims 1 to 16, comprising administering cells.

19. 19. The method of any one of claims 1 to 18, wherein said population of cells and / or progeny cells is autologous or allogeneic and / or the soluble factors are derived from autologous or allogeneic cells.

20. 20. The method of any one of claims 1 to 19, wherein the population of cells and / or their progeny is expanded in culture prior to administration and / or obtaining the soluble factor.

21. 21. The method of any one of claims 1 to 20, wherein the population of mesenchymal progenitor or stem cells express tissue non-specific alkaline phosphatase (TNAP) and / or the progeny cells and / or soluble factors are derived from mesenchymal progenitor or stem cells that express TNAP.

22. The population of mesenchymal progenitor or stem cells is ６ and / or the progeny cells and / or soluble factors express angiopoietin-1 (Ang1) in an amount of at least 0.1 μg per cell. ６ The method according to any one of claims 1 to 21, wherein the cells are derived from mesenchymal precursor or stem cells that express Ang1 in an amount of at least 0.1 µg per cell.

23. The population of mesenchymal progenitor or stem cells is ６ and / or the progeny cells and / or soluble factors express less than about 0.05 μg of vascular endothelial growth factor (VEGF) per cell. ６ 23. The method of any one of claims 1 to 22, wherein the progeny are derived from mesenchymal precursor or stem cells that express VEGF in an amount less than about 0.05 μg per cell.

24. 24. The method of any one of claims 1 to 23, wherein said population of mesenchymal progenitor or stem cells express a ratio of Ang1:VEGF of at least about 2:1 and / or said progeny cells and / or soluble factors are derived from mesenchymal progenitor or stem cells that express a ratio of Ang1:VEGF of at least about 2:

1.

25. The method according to any one of claims 1 to 24, wherein the mesenchymal precursor cells or stem cells and / or their progeny cells and / or soluble factors derived therefrom are administered in the form of a composition comprising the mesenchymal precursor cells or stem cells and / or their progeny cells and / or soluble factors derived therefrom and a carrier and / or excipient.

26. A population of mesenchymal precursor or stem cells and / or their progeny and / or soluble factors derived therefrom for use in treating or preventing progressive heart failure in a subject who has had a myocardial infarction (MI), said subject having proximal left anterior descending (LAD) artery disease.

27. Use of a population of mesenchymal progenitor or stem cells and / or their progeny and / or soluble factors derived therefrom in the manufacture of a medicament for treating or preventing progressive heart failure in a subject who has had a myocardial infarction (MI), wherein the subject has proximal left anterior descending (LAD) artery disease.

28. 28. The population of mesenchymal progenitor or stem cells of claim 26, or the use of claim 27, wherein the subject has persistent left ventricular dysfunction.

29. 29. The population of mesenchymal progenitor or stem cells of claim 26 or 28, or the use of claim 27 or 28, wherein the subject has a LVESV of more than 70 ml.

30. 30. The population of mesenchymal progenitor or stem cells of any one of claims 26, 28 or 29, or the use of any one of claims 27 to 29, wherein the subject has a LVEF of less than about 45%.

31. 30. The population of mesenchymal progenitor or stem cells of any one of claims 26, 28 or 29, or the use of any one of claims 27 to 29, wherein the subject has a LVEF of less than about 40%.

32. The population of mesenchymal precursor or stem cells according to any one of claims 26, 28 to 31 or the use according to any one of claims 27 to 31, wherein the mesenchymal precursor or stem cells and / or their progeny and / or soluble factors derived therefrom are administered between about 1 and 7 days after myocardial infarction.

33. The population of mesenchymal precursor or stem cells according to any one of claims 26, 28 to 31, or the use according to any one of claims 27 to 31, wherein the mesenchymal precursor or stem cells and / or their progeny and / or soluble factors derived therefrom are administered between about 4 and 6 days after myocardial infarction.

34. 34. The population of mesenchymal progenitor or stem cells according to any one of claims 26, 28 to 33, or the use according to any one of claims 27 to 33, wherein the subject has creatine kinase-MB and / or troponin and / or myoglobin greater than about four times the upper limit of normal.

35. The population of mesenchymal precursor cells or stem cells according to any one of claims 26, 28 to 34, or the use according to any one of claims 27 to 34, wherein the subject has an infarct size of between about 10-25% of the left ventricle.

36. The population of mesenchymal precursor cells or stem cells according to any one of claims 26, 28 to 34, or the use according to any one of claims 27 to 34, wherein the subject has an infarct size of more than about 18.5% of the left ventricle.

37. The population of mesenchymal progenitor or stem cells according to any one of claims 26, 28 to 36, or the use according to any one of claims 27 to 36, wherein the LVEF and / or infarct size are measured by cardiovascular magnetic resonance imaging (cMR).

Images