Methods for treating heart failure

Abstract

The present disclosure relates to methods for treating or preventing heart failure in a subject with an elevated left ventricular end-systolic volume (LVESV). In particular, the present disclosure relates to methods for treating or preventing heart failure in a subject with an LVESV greater than 70 ml (>70 ml).

Description

[0001] This application is a divisional application of Chinese invention patent application No. 201580076679.5, whose application date is December 22, 2015 and invention name is “Method for treating heart failure”.

[0002] Citation to Related Applications

[0003] This application claims priority from Australian patent application number 2014905243 filed on 23 December 2015, the entire contents of which are incorporated by reference.

[0004] Public domain

[0005] The present disclosure relates to methods for treating or preventing heart failure in a subject having an elevated left ventricular end-systolic volume (LVESV). Background Art

[0006] Heart failure due to myocardial infarction (MI) remains one of the leading causes of mortality and morbidity in developed countries. A recent update of U.S. Medicare records has been published, evaluating data on 350,509 acute MI hospitalizations involving patients aged >65 years who were discharged alive after their 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 readmitted to the hospital. In the month after 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.

[0007] Patients with larger infarctions and more post-infarction LV dysfunction after MI are at significantly increased risk of experiencing mid- to long-term cardiac events and death. Specifically, anterior wall infarctions, larger infarctions, and more LV dysfunction in the late 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).

[0008] There is a strong need in the art for new and effective methods for preventing and treating heart failure, particularly treatment methods that avoid the need for heart transplants where organ availability is rare. In particular, systems that allow for reversal of heart tissue damage or repair of heart tissue defects without presenting the risks and potential complications associated with general anesthesia and open heart surgery.

[0009] Public Content

[0010] Unexpectedly, preclinical cardiac studies with mesenchymal lineage precursor cells have demonstrated the best results in the context of the largest amount of myocardial injury. This creates the following possibility: signals from damaged myocardium are necessary for the "cross-talk" at the tissue level between the underlying biochemical / physiological disorder and mesenchymal lineage precursor cells to release their "payload", i.e., paracrine factors. This observation has been supported by studies that have demonstrated that there is no significant mesenchymal lineage precursor cell-mediated effect when cells are injected into healthy myocardial tissue.

[0011] The present disclosure is based on the observation that myocardial administration of mesenchymal precursor cells (MPCs) in patients with advanced chronic heart failure due to significant left ventricular contractile abnormalities (i.e., systolic dysfunction) enhances the efficacy signal of MPCs. In particular, the present disclosure is based on the finding that subjects with moderately severe left ventricular contractile impairment respond well to transendocardial delivery of MPCs, and the greater the magnitude of baseline left ventricular contractile abnormalities in patients with chronic heart failure due to left ventricular systolic dysfunction, the greater the benefit of the cardioprotective effect associated with MPCs.

[0012] More specifically, the inventors have discovered that using a baseline left ventricular end-systolic volume (LVESV) cutoff in heart failure subjects identifies a population of subjects with advanced heart failure at high risk for major adverse cardiac events (MACE), and identifies a population of subjects that best responds to MPC administration.

[0013] The present disclosure provides a method for treating or preventing heart failure in a subject, comprising administering to the subject a population of mesenchymal lineage precursor or stem cells and / or progeny thereof and / or soluble factors derived therefrom, wherein the subject has an elevated left ventricular end-systolic volume (LVESV) greater than 70 mL.

[0014] In one example, the elevated LVESV is due to acute myocardial infarction. In another example, the elevated LVESV is due to chronic congestive heart failure.

[0015] In another example, the method comprises the following steps:

[0016] i) selecting subjects with an elevated LVESV greater than 70 mL, and

[0017] ii) administering to the subject a population of mesenchymal lineage precursor or stem cells and / or progeny thereof and / or soluble factors derived therefrom.

[0018] In another example, the method comprises the following steps:

[0019] i) subjects diagnosed with heart failure;

[0020] ii) selecting a cohort of diagnosed subjects with an elevated LVESV greater than 70 mL; and

[0021] iii) administering to the subject a population of mesenchymal lineage stem or precursor cells and / or their progeny and / or soluble factors derived therefrom.

[0022] In one example, the LVESV is greater than 80 mL, greater than 90 mL, greater than 100 mL, greater than 100 mL, greater than 110 mL, greater than 120 mL, or greater than 130 mL. In another example, the LVESV is greater than 80 mL / m 2 , greater than 90mL / m 2 , greater than 100mL / m 2 , greater than 110mL / m 2 , greater than 120mL / m or greater than 130mL / m 2 .

[0023] In one example, the cells and / or their progeny and / or soluble factors are administered to the subject via a catheter-based system. In another example, the cells and / or their progeny and / or soluble factors are administered to the subject's myocardium at or near tissue damage using a catheter inserted into the subject's venous system. In another example, the cells and / or their progeny and / or soluble factors can be administered systemically. Delivery of cells and / or their progeny and / or soluble factors can be performed after identification of a myocardial area in need of treatment.

[0024] In further examples, the cells and / or progeny thereof and / or soluble factors are administered to the subject by transendocardial injection, intracoronary infusion, or transepicardial injection.

[0025] In further examples, the subject is characterized as having a left ventricular ejection fraction (LVEF) of less than or equal to about 35%. In another example, the subject has a LVEF of less than about 35%, less than about 30%, less than about 25%, or less than about 20%.

[0026] In one example, heart failure is due to hypertension, cardiomyopathy (ischemic or non-ischemic), myocarditis, obesity or diabetes. In other examples, mesenchymal lineage stem cells or precursor cells and / or their progeny and / or soluble factors derived therefrom are administered to the subject after diagnosis of heart failure.

[0027] In one example, the subject has acute cardiomyopathy. In another example, the subject has ischemic cardiomyopathy. In another example, the subject has non-ischemic cardiomyopathy. In another example, the subject has dilated or congestive cardiomyopathy. In another example, the subject has restrictive cardiomyopathy.

[0028] In another example, mesenchymal lineage stem cells or precursor cells and / or its offspring and / or soluble factors derived therefrom are applied to the subject after acute myocardial infarction. In another example, mesenchymal lineage stem cells or precursor cells and / or its offspring and / or soluble factors derived therefrom are applied to the subject for about 1-7 days after diagnosing heart failure. In another example, mesenchymal lineage stem cells or precursor cells and / or its offspring and / or soluble factors derived therefrom are applied to the subject for about 1-7 days after myocardial infarction. In another example, after diagnosing heart failure, or about 3-5 days after myocardial infarction, mesenchymal lineage stem cells or precursor cells and / or its offspring and / or soluble factors derived therefrom are applied to the subject.

[0029] In another example, the subject has an infarct size of about 5%-30%, or about 10%-20% of the left ventricle.

[0030] Methods for diagnosing heart failure will be familiar to those skilled in the art. Examples include physical examination of the subject, electrocardiogram, chest x-ray, determination of BNP and / or troponin levels by blood test, echocardiography, Doppler ultrasound, Holter monitor, nuclear heart scan, cardiac catheterization, stress test or cardiac magnetic resonance imaging (cardiac MRI).

[0031] Methods for selecting subjects with an elevated LVESV greater than 70 mL according to the present disclosure are familiar to those skilled in the art. For example, the LVESV of a subject may be measured by equipment known in the art, including but not limited to two-dimensional echocardiography, magnetic resonance tomography, cardiac computed tomography (CT), or biplane left ventriculography. In addition, those skilled in the art will be familiar with the limits and boundaries of the term "elevated LVESV", as that term is understood in the art and as described elsewhere herein.

[0032] The administration of mesenchymal lineage stem cells or precursor cells and / or their offspring according to the present disclosure can be performed on subjects receiving drugs or other treatments for heart failure and / or its symptoms. Alternatively, it can be performed on subjects who have not received other drugs or treatments for heart failure. In some instances, subjects who have previously undergone coronary artery bypass grafting (CABG) or left ventricular assist device (LVAD) implantation are administered with mesenchymal lineage stem cells or precursor cells and / or their offspring according to the present disclosure. The CABG or LVAD implantation regimen experienced by the subject may or may not be accompanied by the simultaneous administration of mesenchymal lineage stem cells or precursor cells and / or their offspring according to the present disclosure. In some instances, individuals who are undergoing CABG or LVAD implantation are administered with mesenchymal lineage stem cells or precursor cells and / or their offspring according to the present disclosure. In other instances, subjects who have not yet received and / or will not receive any therapy for treating damaged / defective cardiac tissue are administered with mesenchymal lineage stem cells or precursor cells and / or their offspring according to the present disclosure.

[0033] The present disclosure also provides methods for treating or preventing heart failure in a heart having an elevated LVESV greater than 70 mL, comprising administering to the heart a population of mesenchymal lineage stem or precursor cells and / or progeny thereof and / or soluble factors derived therefrom.

[0034] In one example, the LVESV is greater than 80 mL, greater than 90 mL, greater than 100 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 , greater than 90mL / m 2 , greater than 100mL / m 2 , greater than 110mL / m 2 or greater than 120 mL / m 2 .

[0035] In another example, the present disclosure provides a method for reducing elevated left ventricular end-systolic volume (LVESV) in a heart having an LVESV greater than 70 mL, comprising administering to the heart a population of mesenchymal lineage stem or precursor cells and / or progeny thereof and / or soluble factors derived therefrom.

[0036] In one example, the LVESV is greater than 80 mL, greater than 90 mL, greater than 100 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 , greater than 90mL / m 2 , greater than 100mL / m 2 , greater than 110mL / m 2or greater than 120 mL / m 2 .

[0037] In another example, the reduction in LVESV is at least 20%, at least 15%, at least 10%, at least 5%, or at least 2% compared to the same heart that has not been administered mesenchymal lineage stem or precursor cells and / or progeny thereof and / or soluble factors derived therefrom.

[0038] In another example, LVESV is measured at about 6 months, about 12 months, about 24 months, or about 36 months after administration of the mesenchymal lineage stem or precursor cells and / or their progeny and / or soluble factors derived therefrom.

[0039] In one example, the cells and / or their progeny and / or soluble factors are administered to the heart via a catheter-based system. In other examples, the cells and / or their progeny and / or soluble factors are administered to the myocardium at or near the site of tissue damage. Delivery of cells and / or their progeny and / or soluble factors may be performed after identification of an area of ​​the myocardium in need of treatment.

[0040] In further examples, the cells and / or progeny thereof and / or soluble factors are administered to the heart by transendocardial injection, intracoronary infusion, or transepicardial injection.

[0041] In additional examples, the heart is further characterized as having a left ventricular ejection fraction (LVEF) of less than about 55%. In additional examples, the heart is further characterized as having a left ventricular ejection fraction (LVEF) of less than or equal to about 35%. In another example, the heart has a LVEF of less than about 35%, less than about 30%, less than about 25%, or less than about 20%.

[0042] In one example, the elevated LVESV is due to acute myocardial infarction. In another example, the elevated LVESV is due to chronic congestive heart failure.

[0043] In another example, mesenchymal lineage stem cells or precursor cells and / or its offspring and / or soluble factors derived therefrom are applied to the heart after diagnosing heart failure. In another example, mesenchymal lineage stem cells or precursor cells and / or its offspring and / or soluble factors derived therefrom are applied to the heart after acute myocardial infarction. In another example, mesenchymal lineage stem cells or precursor cells and / or its offspring and / or soluble factors derived therefrom are applied to the heart for about 1-7 days after diagnosing heart failure. In another example, mesenchymal lineage stem cells or precursor cells and / or its offspring and / or soluble factors derived therefrom are applied to the heart for about 1-7 days after acute myocardial infarction. In another example, mesenchymal lineage stem cells or precursor cells and / or its offspring and / or soluble factors derived therefrom are applied to the heart after diagnosing heart failure or for about 3-5 days after myocardial infarction.

[0044] In another example, the heart has a left ventricular infarct size of about 10%-35%, or 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.

[0045] In some embodiments, the methods of the present disclosure further comprise administering to the heart or to the subject a drug targeting STRO-1. + In another example, the method of the present disclosure comprises administering to the heart or to the subject a protein targeting STRO-1. 亮 Cells are enriched for a population of mesenchymal lineage stem or precursor cells and / or their progeny and / or soluble factors derived therefrom.

[0046] In another example, the population of mesenchymal lineage stem or precursor cells expresses tissue non-specific alkaline phosphatase (TNAP), and / or progeny cells and / or soluble factors are derived from mesenchymal lineage cells expressing TNAP. In another example, the population of mesenchymal lineage stem or precursor cells expresses tissue non-specific alkaline phosphatase (TNAP), and / or progeny cells and / or soluble factors are derived from mesenchymal lineage cells expressing TNAP. 6 The amount of angiopoietin-1 (Ang1) and / or progeny cells and / or soluble factors derived from the culture medium at a concentration of at least 0.1 μg / 10 6 In another example, the population of mesenchymal lineage stem or precursor cells expresses Ang1 at a concentration of at least 0.5 μg / 10 6 In another example, the population of mesenchymal lineage stem or precursor cells expresses Ang1 at a level of at least 1 μg / 10 6 In another example, the population of mesenchymal lineage stem or precursor cells expresses Ang1 at less than about 0.05 μg / 106 The amount of vascular endothelial growth factor (VEGF) and / or progeny cells and / or soluble factors derived from the endothelial growth factor (VEGF) expressed in the endothelial growth factor (VEGF) was less than about 0.05 μg / 10 6 In another example, the population of mesenchymal lineage stem or precursor cells expresses VEGF at less than about 0.03 μg / 10 6 In another example, a population of mesenchymal lineage stem cells or precursor cells expresses VEGF at a ratio of at least about 2: 1: VEGF and / or progeny cells and / or soluble factors are derived from mesenchymal lineage stem cells or precursor cells expressing Ang1: VEGF at a ratio of at least about 2: 1. In another example, a population of mesenchymal lineage stem cells or precursor cells expresses Ang1: VEGF at a ratio of at least about 10: 1: VEGF. In another example, a population of mesenchymal lineage stem cells or precursor cells expresses Ang1: VEGF at a ratio of at least about 20: 1: VEGF. In another example, a population of mesenchymal lineage stem cells or precursor cells expresses Ang1: VEGF at a ratio of at least about 30: 1: VEGF.

[0047] In one example, a population of mesenchymal lineage stem or precursor cells and / or their progeny are administered to the heart or to the subject in a therapeutically effective amount.

[0048] In one example, the population of mesenchymal lineage stem or precursor cells and / or their progeny are administered to the heart or subject in multiple doses. In another example, the methods of the present disclosure include administering 1x10 6 Up to 8x10 8 In another example, the method of the present disclosure comprises administering about 1.5 x 10 8 cells.

[0049] In one example, mesenchymal lineage stem cells or precursor cells and / or their progeny cells and / or soluble factors derived therefrom are administered in the form of a composition comprising mesenchymal lineage stem cells or precursor cells and / or their progeny cells and / or soluble factors derived therefrom and a pharmaceutically acceptable carrier and / or excipient. In another example, a population of mesenchymal lineage stem cells or precursor cells and / or their progeny have been cultured and expanded prior to administration and / or prior to obtaining the soluble factors.

[0050] The present disclosure also provides for the use of a population of mesenchymal lineage stem or precursor cells and / or progeny thereof and / or soluble factors derived therefrom or a composition described herein for treating heart failure in a subject with an elevated LVESV greater than 70 mL.

[0051] The present disclosure also provides for the use of a population of mesenchymal lineage stem or precursor cells and / or progeny thereof and / or soluble factors derived therefrom or a composition described herein for treating a heart having an elevated LVESV greater than 70 mL.

[0052] In one example according to any use described herein, the elevated LVESV is due to acute myocardial infarction. In another example according to any use described herein, the elevated LVESV is due to chronic congestive heart failure.

[0053] The present disclosure also provides a population of mesenchymal lineage stem or precursor cells and / or progeny thereof and / or soluble factors derived therefrom for use in the manufacture of a medicament for treating heart failure in a subject with an elevated left ventricular end-systolic volume (LVESV) greater than 70 mL.

[0054] The present disclosure also provides a population of mesenchymal lineage stem or precursor cells and / or progeny thereof and / or soluble factors derived therefrom for use in the manufacture of a medicament for treating a heart having an elevated left ventricular end-systolic volume (LVESV) greater than 70 mL.

[0055] The present disclosure also provides a population of mesenchymal lineage stem or precursor cells and / or progeny thereof and / or soluble factors derived therefrom or a composition described herein for use in treating heart failure in a subject with an elevated LVESV greater than 70 mL.

[0056] The present disclosure also provides a population of mesenchymal lineage stem or precursor cells and / or progeny thereof and / or soluble factors derived therefrom or a composition described herein for use in treating a heart having an elevated left ventricular end-systolic volume (LVESV) greater than 70 mL.

[0057] In another example, the present disclosure provides use of a population of mesenchymal lineage stem or precursor cells and / or progeny thereof and / or soluble factors derived therefrom or a composition described herein for reducing left ventricular end-systolic volume (LVESV) in a heart with an elevated LVESV greater than 70 mL.

[0058] In another example, the present disclosure provides a population of mesenchymal lineage stem or precursor cells and / or progeny thereof and / or soluble factors derived therefrom or a composition described herein for use in reducing left ventricular end-systolic volume (LVESV) in a heart with an elevated LVESV greater than 70 mL.

[0059] In one example according to any population or use described herein, 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, LVESV is greater than 80 mL / m 2 , greater than 90mL / m 2 , greater than 100mL / m 2 , greater than 110mL / m 2 , or greater than 120 mL / m 2 .

[0060] In further examples according to any of the methods or uses described herein, a population of mesenchymal lineage precursor cells and / or their progeny are isolated or purified.

[0061] In other examples, the colony of mesenchymal lineage stem cell or precursor cell and / or its offspring are derived from donor subjects.Donor subjects can be the same subjects that accept cells and / or their offspring and / or soluble factors derived therefrom are used, in which case cells are autologous.In another example, donor subjects are different subjects that accept cells and / or their offspring and / or soluble factors derived therefrom are used, in which case cells are allogenic.

[0062] In a further example, the subject has a left ventricular ejection fraction (LVEF) of less than about 55%. In another example according to any use described herein, the subject has a LVEF of less than or equal to about 35%. In another example, the subject has a LVEF of less than about 35%, less than about 30%, less than about 25%, or less than about 20%.

[0063] In another example according to any use described herein, the heart has a left ventricular infarct size of about 5%-30%, or about 10%-20%. In another example according to any use described herein, the subject has a left ventricular infarct size of about 5%-30%, or about 10%-20%.

[0064] In another example, the present disclosure provides a population of mesenchymal stem cells or lineage precursor cells and / or progeny thereof and / or soluble factors derived therefrom for use in the manufacture of a medicament for reducing left ventricular end-systolic volume (LVESV) in a heart with an elevated LVESV greater than 70 mL.

[0065] The present disclosure also provides a kit, which includes a population of mesenchymal lineage stem or precursor cells and / or their progeny and / or soluble factors derived therefrom or a composition described herein and a delivery device for administering the cells, and / or their progeny and / or soluble factors. In one example, the delivery device is a catheter.

[0066] In another example, the subject according to the present disclosure is a mammal. In another example, the subject is a human, including a teenager or a child. In a specific example, the subject is greater than or equal to 18 years old.

[0067] In further examples, the subject has had an episode of heart failure within 12 months prior to administration of the population of mesenchymal lineage stem or precursor cells and / or progeny thereof and / or soluble factors derived therefrom.

[0068] In another example, the subject is classified as New York Heart Association (NYHA) class II, III or IV. Class II is defined as mild, in which the subject experiences fatigue and shortness of breath during moderate physical activity. Class II is defined as moderate, in which the patient experiences shortness of breath even during mild physical activity. Class IV or terminal positioning is severe, in which the patient is exhausted even at rest.

[0069] In another example, the subject has a reduced baseline 6-minute walk test (6MWT), meaning that the distance the subject travels in a 6-minute period is reduced relative to non-heart failure subjects.

[0070] In another example, the subject has a left ventricular end-diastolic volume (LVEDS) greater than 150 mL. In another example, the subject has a LVESV greater than 170 mL.

[0071] In another example, the subject is also co-administered with conventional heart failure medications. In other examples, heart failure medications include, but are not limited to, one or more of the following: beta blockers, ACE inhibitors, or angiotensin receptor blockers.

[0072] In another example, the subject has a higher major adverse cardiac event (MACE) rate. In a further example, the subject has a>50% heart failure-MACE (HF-MACE) rate within 36 months. In another example, the subject has a>60% HF-MACE rate, or>65% HF-MACE rate, or>70% HF-MACE rate within 36 months. BRIEF DESCRIPTION OF THE DRAWINGS

[0073] Figure 1 A dose-dependent effect of MPC on cardiac remodeling based on left ventricular (LV) volume was demonstrated.

[0074] Figure 2 Shown is the HF-MAC obtained by Kaplan-Meier curve 36 months after treatment with 150 million MPCs.

[0075] Figure 3-1Shown are baseline left ventricular end-systolic volume (LVESV) values ​​in subjects administered placebo (Control) and mesenchymal precursor cells (MPCs) (150 million cells) stratified by LVESV less than or equal to 100 mL or greater than 100 mL.

[0076] Figure 3-2 Shown are the changes in LVESV values ​​at 6 months in subjects administered placebo (control) and MPC (150 million cells) stratified by LVESV less than or equal to 100 mL or greater than 100 mL.

[0077] Figure 4-1 Shown are baseline left ventricular end-systolic volume (LVEDV) values ​​in subjects administered placebo (control) and MPC (150 million cells) stratified by LVESV less than or equal to 100 mL or greater than 100 mL.

[0078] Figure 4-2 Shown are changes in LVEDV values ​​at 6 months in subjects administered placebo (control) and MPC (150 million cells) stratified by LVESV less than or equal to 100 mL or greater than 100 mL.

[0079] Figure 5-1 Shown are baseline left ventricular ejection fraction (LVEF) values ​​in subjects administered placebo (control) and MPC (150 million cells) stratified by LVESV less than or equal to 100 mL or greater than 100 mL.

[0080] Figure 5-2 Shown are changes in LVEF values ​​at 6 months in subjects administered placebo (control) and MPC (150 million cells) stratified by LVESV less than or equal to 100 mL or greater than 100 mL.

[0081] Figure 6 Shown are the changes in LVESV values ​​at 6 months in subjects administered placebo (control) and MPC (150 million cells) stratified by LVESV greater than 70 mL.

[0082] Figure 7 Shown are the changes in LVESV values ​​at 6 months in subjects administered placebo (control) and MPC (150 million cells) stratified by LVESV greater than 80 mL.

[0083] Figure 8 Shown are the changes in LVESV values ​​at 6 months in subjects administered placebo (control) and MPC (150 million cells) stratified by LVESV greater than 90 mL.

[0084] Fig. 9 Shown are the changes in LVESV values ​​at 6 months in subjects administered placebo (control) and MPC (150 million cells) stratified by LVESV greater than 100 mL.

[0085] Fig.10 Shown are the changes in LVESV values ​​at 6 months in subjects administered placebo (control) and MPC (150 million cells) stratified by LVESV greater than 110 mL.

[0086] Fig.11 Shown are the changes in LVESV values ​​at 6 months in subjects administered placebo (control) and MPC (150 million cells) stratified by LVESV greater than 120 mL.

[0087] Fig.12 Kaplan-Meier curves of MF-MACE at 36 months after treatment in patients with LVESV>100 ml are shown. DETAILED DESCRIPTION

[0088] Throughout this specification, references to individual steps, compositions of matter, groups of steps or groups of compositions of matter should be understood to cover both one and multiple (i.e. one or more) of those steps, compositions of matter, groups of steps or groups of compositions of matter, unless expressly stated otherwise or the context requires otherwise.

[0089] Those skilled in the art will appreciate that, in addition to those specifically described, the disclosure described herein is susceptible to changes and modifications. It should be understood that the disclosure includes all such changes and modifications. The disclosure also includes all steps, features, compositions and compounds cited or indicated in this specification individually or collectively, and any and all combinations or any two or more of the steps or features.

[0090] The present disclosure is not to be limited in scope by the specific embodiments described herein, which are intended for purposes of example only. As described herein, functionally equivalent products, compositions, and methods are clearly within the scope of the present disclosure.

[0091] Unless expressly stated otherwise, any example disclosed herein should be considered applicable mutatis mutandis to any other example.

[0092] Unless specifically defined otherwise, all technical and scientific terms used herein should be interpreted as having the same meaning as commonly understood by one of ordinary skill in the art (e.g., in cell culture, molecular genetics, stem cell differentiation, immunology, immunohistochemistry, protein chemistry, and biochemistry).

[0093] Unless otherwise indicated, the surgical techniques used in this disclosure are standard procedures well known to those skilled in the art.

[0094] Methods for obtaining and enriching populations of mesenchymal lineage stem or precursor cells are known in the art. For example, enriched populations of mesenchymal lineage stem or precursor cells can be obtained by using flow cytometry and cell sorting procedures based on the use of cell surface markers expressed on mesenchymal lineage stem or precursor cells.

[0095] All documents cited or referred to herein and all documents cited or referred to in documents cited herein, together with any manufacturer's instructions, descriptions, product specifications and product sheets for any products mentioned herein, or any documents incorporated herein by reference, are hereby incorporated by reference in their entirety.

[0096] Definition of Choice

[0097] The term "and / or", such as "X and / or Y", should be understood as "X and Y" or "X or Y", and should be considered to provide clear support for both or either meanings.

[0098] As used herein, unless otherwise indicated, the term "about" means + / - 10%, more preferably + / - 5%, of the specified value.

[0099] Throughout the specification, the word "comprises / comprising" or variations will be understood to imply the inclusion of stated elements, integers or steps, or groups of elements, integers or steps, but not the exclusion of any other elements, integers or steps, or groups of elements, integers or steps.

[0100] As used herein, the singular form "a", "an", and "the" include singular and plural references unless the context dictates otherwise.

[0101] As used herein, the term "heart failure" can be used interchangeably with the term "congestive heart failure (CHF)" and refers to a condition in which the heart cannot pump enough blood to other organs of the body due to, for example, myocardial dysfunction, myocardial weakness (known as "cardiomyopathy"), and other myocardial-related reasons. Congestive heart failure is characterized by left ventricular (LV) chamber dilation, decreased LV contractility, and elevated circulating catecholamine levels, among other effects. In another example, heart failure occurs due to ischemia and other reperfusion and other non-ischemic factors. Heart failure includes, but is not limited to, the following symptoms or signs, alone or in combination: cardiac reperfusion injury, compensatory hypertrophy, end-stage heart failure in humans, hypertensive cardiomyopathy, left ventricular hypertension, left or right ventricular dilation, left or right ventricular failure, maladaptive hypertrophy, myocardial structural disorder (apoptosis and loss of myocardial cells) and myocardial dysfunction (loss of contraction and / or relaxation) and a heart with pressure overload.

[0102] By "isolated" or "purified" is meant a cell that has been separated from at least some components of its natural environment. The term includes gross physical separation of a cell from its natural environment (e.g., removal from a donor). The term "isolated" includes altering the relationship of a cell to its immediate neighbors, such as by dissociation. The term "isolated" does not refer to cells in a tissue section. When used to refer to a population of cells, the term "isolated" includes a population of cells resulting from the proliferation of an isolated cell of the present disclosure.

[0103] As used herein, the term "left ventricular hypertension (LVH)" is a condition in which the heart muscle responds to increased circulatory resistance by becoming enlarged. However, over time, the fibers of the hypertrophic heart muscle thicken and shorten and are therefore less able to relax. Hypertension causes the heart muscle to work harder. The resulting hypertrophy is a product of the thickening or shortening of the heart's muscle fibers. Under these conditions, it becomes more difficult for the heart to relax and go through the normal cycle of contraction and relaxation. Myocardial changes occur in collagen, resulting in increased stiffness. The result of this process is a heart that is less able to meet the output demands of normal circulation.

[0104] As used herein, the term "left ventricular dilatation" refers to an enlargement of the left ventricle, which can increase the volume of blood ejected from the ventricle, temporarily improving cardiac output. However, the increased size of the ventricle also leads to a decrease in the percentage of left ventricular volume of blood incurred (called ejection fraction), and has significant physiological significance. Left ventricular dilatation is a recognized precursor and sign of cardiac dysfunction and congestive heart failure after myocardial infarction. Similarly, right ventricular dilatation refers to right ventricular enlargement and related signs or symptoms.

[0105] As used herein, the term "left ventricular failure" refers to a condition in which the left side of the heart cannot pump blood effectively. This results in reverse flow of blood into the lungs, pressure, and / or congestion. Signs indicative of left ventricular failure include a laterally displaced apex beat. A gallop rhythm may be heard as a sign of increased blood flow or increased pressure within the heart.

[0106] As used herein, the term "cardiomyopathy" refers to a condition in which the heart muscle (myocardium) becomes inflamed and enlarged. Several different types of cardiomyopathy are known in the art, including dilated cardiomyopathy, in which the heart muscle is stretched and thinned, hypertrophic cardiomyopathy, in which the heart muscle cells enlarge and cause the heart wall to thicken, and restrictive cardiomyopathy, in which the heart becomes stiff and rigid due to abnormal tissue (e.g., scar tissue).

[0107] As used herein, the term "myocardial infarction" is also understood to refer to a heart attack. A heart attack occurs when blood stops flowing normally to a part of the heart and the heart muscle is damaged because it does not receive enough oxygen. This can occur when a blockage occurs in one of the coronary arteries that supply blood to the heart.

[0108] As used herein, the term "left ventricular end-systolic volume" (LVESV) refers to the volume of blood in the left ventricle at the end of contraction, or systole, and the beginning of filling (filing), or diastole. It refers to the lowest blood volume in the ventricle at any point in the cardiac cycle. Normal values ​​for men typically range from 22-58 mL and for women from 19-49 mL. For explanation, LVESV values ​​herein refer to baseline LVESV values, i.e., LVESV values ​​determined before administering mesenchymal lineage precursor cells or stem cells and / or their progeny and / or soluble factors derived therefrom to a subject.

[0109] The term "elevated LVESV" will be understood by those skilled in the art. Generally, it will be understood to mean a value above the normal LVESV range for males or females. In one example, an elevated LVESV will be understood to mean a value greater than 70 mL. In another example, it will be understood to mean an LVESV value greater than 100 ml.

[0110] As used herein, the term "left ventricular end-systolic volume index" (LVESVI) refers to the indexed left ventricular end-systolic volume. This value is usually expressed as mL / m 2 The terms LVESV and LVESVI may be used interchangeably herein.

[0111] As used herein, the term "left ventricular ejection fraction" (LVEF) refers to how well the heart pumps with each contraction. The ejection fraction (EF) is usually expressed as a percentage, which is the percentage of blood that leaves the heart with each contraction. A normal LVEF ranges from 55-70%. An LVEF of 65%, for example, means that 65% of the total amount of blood in the left ventricle is pumped out with each heartbeat. Since the left ventricle is the main pumping chamber of the heart, EF is usually measured only in the left ventricle (LV). An LVEF of more than 55% is considered normal. An LVEF of less than 50% is considered reduced. Experts vary in their opinions about an EF between 50% and 55%, and some consider this to be a borderline range.

[0112] As used herein, the term "left ventricular end-diastolic volume" (LVEDV) refers to the volume of blood in the right ventricle at the end of contraction, or systole, and the beginning of filling, or diastole. It refers to the lowest blood volume in the ventricle at any point in the cardiac cycle. Typically, the LVEDV of a normal healthy subject is about 120 mL. For explanation, LVEDV values ​​herein refer to baseline LVEDV values, i.e., LVEDV values ​​determined before administering mesenchymal lineage precursor cells and / or their progeny and / or soluble factors derived therefrom to a subject.

[0113] As used herein, the term "treating", "treat" or "treatment" includes administering a population of mesenchymal lineage stem cells or precursor cells and / or their progeny and / or soluble factors derived therefrom, thereby reducing or eliminating at least one symptom of heart failure. In a specific example, compared to a baseline value (i.e., before administering a population of mesenchymal lineage stem cells or precursor cells and / or their progeny and / or soluble factors derived therefrom), treatment reduces the LVESV value by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50%.

[0114] As used herein, the terms "prevent" or "preventing" include administering a population of mesenchymal lineage stem or precursor cells and / or their progeny and / or soluble factors derived therefrom to halt or impede the development of at least one symptom of heart failure.

[0115] As used herein, the term "subject" refers to mammals, including but not limited to mice, rats, apes, humans, domestic animals, and farm animals.

[0116] Mesenchymal lineage precursor cells

[0117] As used herein, the term "mesenchymal lineage precursor cell or stem cell" refers to an undifferentiated multipotent cell with the ability to self-renew while maintaining pluripotency and differentiation into a variety of cell types of mesenchymal origin (e.g., osteoblasts, chondrocytes, adipocytes, stromal cells, fibroblasts, and tendons), or non-mesoderm origin (e.g., hepatocytes, neural cells, and epithelial cells). For the avoidance of doubt, "mesenchymal lineage precursor cells" refer to cells that can differentiate into mesenchymal cells such as bone, cartilage, muscle, and adipocytes, as well as fibrous connective tissue.

[0118] The term "mesenchymal lineage precursor or stem cell" includes both the parent cell and its undifferentiated progeny. The term also includes mesenchymal precursor cells, multipotent stromal cells, mesenchymal stem cells (MSC), perivascular mesenchymal precursor cells and their undifferentiated progeny.

[0119] Mesenchymal lineage precursor cells or stem cells can be autologous, xenogeneic, syngeneic or isogenic. Autologous cells are isolated from the same individual into which they will 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 (e.g., twins, clones, or highly inbred research animal models).

[0120] Mesenchymal lineage precursors or stem cells reside primarily in the bone marrow but have also been shown to be present in a variety of host tissues including, for example, umbilical cord blood and cord, adult peripheral blood, adipose tissue, trabecular bone, and dental pulp.

[0121] In one example, the mesenchymal lineage precursor cell is a STRO-1+ mesenchymal precursor cell (MPC). As used herein, the phrase "STRO-1+ multipotent cell" should be taken to refer to a STRO-1+ and / or TNAP+ progenitor cell capable of forming a multipotent cell colony.

[0122] STRO-1+ pluripotent cells are found in bone marrow, blood, dental pulp cells, adipose tissue, skin, spleen, pancreas, brain, kidney, liver, heart, retina, brain, hair follicles, intestines, lungs, lymph nodes, thymus, bones, ligaments, tendons, skeletal muscle, dermis and periosteum; and are capable of differentiating into germline cells, such as mesoderm and / or endoderm and / or ectoderm. Therefore, STRO-1+ pluripotent cells are capable of differentiating into a large number of cell types, including but not limited to adipose tissue, bone tissue, cartilage tissue, elastic tissue, muscle tissue and fibrous connective tissue. The specific lineage orientation and differentiation pathways that these cells enter depend on the various influences of institutional influences and / or endogenous bioactive factors, such as growth factors, cytokines and / or local microenvironment conditions established by the host tissue.

[0123] Mesenchymal lineage precursor cells or stem cells can be isolated from host tissue and enriched by selecting STRO-1+ cells. For example, a bone marrow aspirate from a subject can be further treated with antibodies against STRO-1 or TNAP to enable selection of mesenchymal lineage precursor cells or stem cells. In one example, mesenchymal lineage precursor cells or stem cells can be enriched using the STRO-1 antibody described in (Simmons & Torok-Storb, 1991).

[0124] The term "enriched" or variations thereof are used herein to describe a cell population in which the proportion of a particular cell type or the proportion of multiple specific cell types is increased when compared to an untreated cell population (e.g., cells in its 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 "cell population enriched for STRO-1+ cells" should be considered to provide clear support for the term "cell population containing X% STRO-1+ cells", where X% is a percentage as described herein. In some instances, STRO-1+ cells can form clonogenic colonies, for example, CFU-F (fibroblasts) or a subset thereof (e.g., 50% or 60% or 70% or 70% or 90% or 95%) can have such activity.

[0125] In one example, a cell population is enriched from a cell preparation comprising STRO-1+ cells in a selectable form. In this regard, the term "selectable form" will be understood to mean that the cell expresses a marker (e.g., a cell surface marker) that allows STRO-1+ cell selection. The marker may be STRO-1, but this is not required. For example, as described and / or exemplified herein, cells (e.g., mesenchymal precursor cells) expressing 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 may be STRO-1 bright). Therefore, the indication that a cell is STRO-1+ does not mean that cells are selected only by STRO-1 expression. In one example, cells are selected based on at least STRO-3 expression, for example, they are STRO-3+ (TNAP+).

[0126] Reference to the selection of cells or their populations 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. That is, in some instances, these terms provide support for selection from tissues containing STRO-1+ cells (e.g., mesenchymal precursor cells) or vascularized tissues or tissues containing pericytes (e.g., STRO-1+ pericytes) or any one or more of the tissues described herein.

[0127] In one example, the cells used in the present disclosure express one or more markers selected alone or collectively from the group consisting of TNAP+, VCAM-1+, THY-1+, STRO-2+, STRO-4+ (HSP-90β), CD45+, CD146+, 3G5+, or any combination thereof.

[0128] "Individually" means that the disclosure individually encompasses the marker or marker group, and although an individual marker or marker group may not be individually listed herein, the appended claims may separately and separably define such marker or marker group from each other.

[0129] "Collectively" means that the present disclosure encompasses any number or combination of the markers or marker groups, and although such number or combination of markers or marker groups may not be explicitly listed herein, the appended claims may define such combinations or subcombinations separately and separably from any other combination of markers or marker groups.

[0130] In one embodiment, STRO-1+ cells are STRO-1 亮 (Synonym STRO-1 bri ). In another example, relative to STRO-1 暗 or STRO-1 中间 Cells preferentially enrich for STRO-1 bri cell.

[0131] In another embodiment, STRO-1 bri The cells are additionally one or more of TNAP+, VCAM-1+, THY-1+, STRO-2+, STRO-4+ (HSP-90β) and / or CD146+. For example, the cells are selected for one or more of the aforementioned markers and / or displayed to express one or more of the aforementioned markers. In this regard, cells that display expression of a marker do not need to be explicitly tested, but rather previously enriched or isolated cells can be tested and subsequently used, and the isolated or enriched cells can reasonably be assumed to also express the same marker.

[0132] In one example, the mesenchymal precursor cell (MPC) is a perivascular mesenchymal precursor cell as defined in WO 2004 / 85630, characterized by the presence of the perivascular marker 3G5. For example, the MPC expresses markers of perivascular cells, such as the cell is STRO-1+ or STRO-1 bri In one example, the cell is or was previously or is a cell isolated from a vascularized tissue or organ or a part thereof or is a descendant of such a cell.

[0133] Cells that are called "positive" for a given marker may express low (lo or dark) or high (bright, bri) levels of the marker, depending on the extent to which the marker is present on the cell surface, where the term is related to the fluorescence intensity or other markers used in the cell sorting process. The distinction between lo (or dark or dim) and bri will be understood in the context of the markers used on the specific cell population being sorted. Cells that are called "negative" for a given marker are not necessarily completely absent from the cell. The term means that the marker is expressed by the cell at a relatively very low level, and it produces a very low signal when detectably labeled or is undetectable above background levels, such as the level detected using an isotype control antibody.

[0134] As used herein, the term "bright" or "bri" refers to a marker on the surface of a cell that produces a relatively high signal when detectably labeled. While not wishing to be bound by theory, it is proposed that "bright" cells express more of the target marker protein (e.g., an antigen recognized by STRO-1) than other cells in the sample. For example, when labeled with a FITC-conjugated STRO-1 antibody, STRO-1 is expressed more than non-bright cells (STRO-1 dim / dark) as determined by fluorescence activated cell sorting (FACS) analysis. bri In one example, the "bright" cells make up at least about 0.1% of the brightest labeled bone marrow mononuclear cells contained in the starting sample. In other examples, the "bright" cells make up at least about 0.5%, at least about 1%, at least about 1.5%, or at least about 2% of the brightest labeled bone marrow mononuclear cells contained in the starting sample. In one example, the STRO-1 bright cells are expressed relative to the "background," i.e., STRO-1. - cells, have 2 logs higher expression of STRO-1 surface expression. In contrast, STRO-1 dim and / or STRO-1 intermediate cells have less than 2 logs higher expression of STRO-1 surface expression than "background", typically about 1 log or less.

[0135] As used herein, the term "TNAP" is intended to encompass all isoforms of tissue nonspecific alkaline phosphatase. For example, the term includes liver isoform (LAP), bone isoform (BAP), and kidney isoform (KAP). In one example, TNAP is BAP. In one example, as used herein, TNAP refers to a molecule that can bind to the STRO-3 antibody produced by the hybridoma cell line deposited with the ATCC under the deposit accession number PTA-7282 on December 19, 2005 in accordance with the provisions of the Budapest Treaty.

[0136] Furthermore, in one example, STRO-1+ cells were able to generate clonogenic CFU-F.

[0137] In one example, a considerable proportion of STRO-1+ cells are able to differentiate into at least two different germlineages. Non-limiting examples of lineages that can direct STRO-1+ cells include bone precursor cells; hepatocyte progenitor cells, which are multipotent for bile duct epithelial cells and hepatocytes; neural restricted cells, which can produce glial cell precursors that progress to oligodendrocytes and astrocytes; neuronal precursors that progress to neurons; precursors of myocardium and myocardial cells, glucose-responsive insulin-secreting pancreatic β cell lines. Other lineages include, but are not limited to, odontoblasts, cells that produce dentin, and chondrocytes, as well as the following precursor cells: retinal pigment epithelial cells, fibroblasts, skin cells such as keratinocytes, dendritic cells, hair follicle cells, renal tubular epithelial cells, smooth and skeletal muscle cells, testicular progenitor cells, vascular endothelial cells, tendons, ligaments, cartilage, adipocytes, fibroblasts, bone marrow stroma, myocardium, smooth muscle, skeletal muscle, pericytes, blood vessels, epithelial cells, glial cells, neurons, astrocytes, and oligodendrocytes.

[0138] In one example of the present disclosure, the mesenchymal lineage precursor cell or stem cell is a mesenchymal stem cell (MSC). MSC can be a homogenous composition, or can be a mixed cell population rich in MSC. Homogeneous MSC compositions can be obtained by culturing adherent bone marrow or periosteal cells, and MSC can be identified by specific cell surface markers identified with unique monoclonal antibodies. Methods for obtaining cell populations enriched in MSC are described in, for example, U.S. Patent No. 5,486,359. Alternative sources of MSC include, but are not limited to, blood, skin, umbilical cord blood, muscle, fat, bone, and perichondrium.

[0139] In another example, the mesenchymal lineage precursor or stem cell is a CD29+, CD54+, CD73+, CD90+, CD102+, CD105+, CD106+, CD166+, MHC1+ MSC (e.g., remestemcel-L).

[0140] The isolated or enriched mesenchymal lineage precursor cells or stem cells can be expanded in vitro by culture. The isolated or enriched mesenchymal lineage precursor cells or stem cells can be cryopreserved, thawed, and subsequently expanded in vitro by culture.

[0141] In one example, isolated or enriched mesenchymal lineage precursor cells or stem cells are cultured at 50,000 viable cells / cm 2 Seed in culture medium (serum-free or serum-supplemented), such as α-minimal essential medium (αMEM) supplemented with 5% fetal bovine serum (FBS) and glutamine, and allowed to adhere to the culture vessel overnight at 37° C., 20% O 2. The culture medium is then replaced and / or changed as needed, and the cells are cultured for another 68 to 72 hours at 37° C., 5% O 2 .

[0142] As will be appreciated by those skilled in the art, the mesenchymal lineage precursor cells or stem cells cultured are different from the in vivo cell phenotype. For example, in one embodiment, they express one or more of the following markers: CD44, NG2, DC146 and CD140b. The mesenchymal lineage precursor cells or stem cells cultured are biologically different from in vivo cells, having a higher proliferation rate compared to most non-circulating (quiescent) cells in vivo.

[0143] Mesenchymal lineage precursor cells or stem cells may also be cryopreserved prior to administration to a subject.

[0144] Cell modification

[0145] In one example, the mesenchymal lineage precursor cells or stem cells of the present disclosure may be genetically modified or genetically unmodified and are expressed at a concentration of at least 0.1 μg / 10 6 However, in various embodiments, it is contemplated that the mesenchymal stem cells or precursor cells of the present disclosure may express Ang1 at a level of at least 0.2 μg / 10 6 cells, 0.3 μg / 10 6 cells, 0.4 μg / 10 6 cells, 0.5 μg / 10 6 cells, 0.6 μg / 10 6 cells, 0.7 μg / 10 6 cells, 0.8 μg / 10 6 cells, 0.9 μg / 10 6 cells, 1μg / 10 6 cells, 1.1 μg / 10 6 cells, 1.2 μg / 10 6 cells, 1.3 μg / 10 6 cells, 1.4 μg / 106 cells, 1.5 μg / 10 6 The number of cells expressing Ang1.

[0146] In one example, the mesenchymal lineage precursor or stem cells of the present disclosure are genetically unmodified and are expressed at a concentration of at least 0.1 μg / 10 6 However, in various embodiments of the present invention, it is contemplated that the mesenchymal lineage precursor cells or stem cells may express Ang1 at a concentration of at least 0.2 μg / 10 6 cells, 0.3 μg / 10 6 cells, 0.4 μg / 10 6 cells, 0.5 μg / 10 6 cells, 0.6 μg / 10 6 cells, 0.7 μg / 10 6 cells, 0.8 μg / 10 6 cells, 0.9 μg / 10 6 cells, 1μg / 10 6 cells, 1.1 μg / 10 6 cells, 1.2 μg / 10 6 cells, 1.3 μg / 10 6 cells, 1.4 μg / 10 6 cells, 1.5 μg / 10 6 The number of cells expressing Ang1.

[0147] In another aspect, the mesenchymal lineage precursor cells or stem cells of the present disclosure are expressed at less than about 0.05 μg / 10 6 However, in various embodiments, it is contemplated that the mesenchymal lineage stem cells or precursor cells of the present disclosure may express VEGF at less than about 0.05 μg / 10 6 cells, 0.04μg / 10 6 cells, 0.03μg / 10 6 cells, 0.02μg / 10 6 cells, 0.01 μg / 10 6 cells, 0.009μg / 10 6 cells, 0.008 μg / 10 6 cells, 0.007 μg / 10 6 cells, 0.006μg / 10 6 cells, 0.005 μg / 10 6 cells, 0.004 μg / 10 6 cells, 0.003μg / 10 6 cells, 0.002μg / 10 6cells, 0.001μg / 10 6 The number of cells expressing VEGF.

[0148] In one example, the mesenchymal lineage precursor or stem cells of the present disclosure are genetically unmodified and expressed at less than about 0.05 μg / 10 6 However, in various embodiments of this example, it is contemplated that the mesenchymal lineage precursor cells or stem cells of the present disclosure may express VEGF at less than about 0.05 μg / 10 6 cells, 0.04μg / 10 6 cells, 0.03μg / 10 6 cells, 0.02μg / 10 6 cells, 0.01 μg / 10 6 cells, 0.009μg / 10 6 cells, 0.008 μg / 10 6 cells, 0.007 μg / 10 6 cells, 0.006μg / 10 6 cells, 0.005 μg / 10 6 cells, 0.004 μg / 10 6 cells, 0.003μg / 10 6 cells, 0.002μg / 10 6 cells, 0.001μ / 10 6 The number of cells expressing VEGF.

[0149] The amount of cellular Ang1 and / or VEGF expressed in a composition or culture of mesenchymal lineage precursor cells or stem cells can be determined by methods known to those skilled in the art. Such methods include, but are not limited to, quantitative assays, such as quantitative ELISA assays, for example, or fluorescence-linked immunosorbent assays (FLISA), Western blots, competitive assays, radioimmunoassays, lateral flow immunoassays, flow-through immunoassays, electrochemiluminescence assays, nephelometric-based assays, turbidometric-based assays, fluorescence-activated cell sorting (FACS)-based assays (for detecting Ang-1 or VEGF in culture media of mesenchymal lineage precursor cells or stem cells), surface plasmon resonance (SPR or Biacore).

[0150] However, it should be understood that the scope of the present disclosure is not limited to any specific method for determining the amount or level of Ang1 or VEGF expressed in the mesenchymal lineage precursor cells or stem cells of the present disclosure.

[0151] In one example, the level of Ang1 or VEGF expressed by a composition or culture of a mesenchymal lineage precursor cell or stem cell is determined by an ELISA assay. In such an assay, a cell lysate from a culture of a mesenchymal lineage precursor cell or stem cell is added to the wells of an ELISA plate. The wells can be coated with a primary antibody (monoclonal or polyclonal antibody) for Ang1 or VEGF. Then, the wells are washed and then contacted with a secondary antibody (monoclonal or polyclonal antibody) for the primary antibody. For example, the secondary antibody is conjugated with an appropriate enzyme, such as horseradish peroxidase. The wells can be incubated and then washed after the incubation period. Then, the wells are contacted with a substrate suitable for the enzyme conjugated with the secondary antibody, such as one or more chromogens. The chromogens that can be used include, but are not limited to, hydrogen peroxide and tetramethylbenzidine. After adding the substrate, the wells are incubated for an appropriate period of time. After the incubation is complete, a "stop" solution is added to the wells to stop the reaction of the enzyme with the substrate. The optical density (OD) of the sample is then measured. The optical density of the sample is correlated to the optical density of samples containing known amounts of Ang1 or VEGF to determine the amount of Ang1 or VEGF expressed by the culture of mesenchymal lineage precursor cells or stem cells being tested.

[0152] In another aspect, the mesenchymal lineage precursor cells or stem cells of the present disclosure express Ang1:VEGF at a ratio of at least about 2: 1. However, in various embodiments, it is contemplated that the mesenchymal lineage precursor cells or stem cells of the present disclosure may express Ang1:VEGF at 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.

[0153] In one example, the mesenchymal lineage precursor cells or stem cells of the present disclosure are genetically unmodified and express Ang1:VEGF at a ratio of at least about 2: 1. However, in various embodiments, it is contemplated that the mesenchymal lineage precursor cells or stem cells of the present disclosure may express Ang1:VEGF at 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.

[0154] Methods for determining the Ang1:VEGF expression ratio will be apparent to those skilled in the art. In an example of a method for determining the Ang1 and VEGF expression ratio, the Ang1 and VEGF expression levels are quantified by quantitative ELISA as described above. In such examples, after quantifying the levels of Ang1 and VEGF, the ratio based on the quantitative levels of Ang1 and VEGF can be expressed as: (Ang1 level / VEGF level) = Ang1:VEGF ratio.

[0155] The mesenchymal lineage precursor cells or stem cells disclosed herein can be altered in such a way that upon administration, cell lysis is inhibited. The alteration of antigens can induce immunological non-responsiveness or tolerance, thereby preventing the induction of the effector phase of the immune response (e.g., cytotoxic T cell production, antibody production, etc.) that ultimately causes foreign cell rejection in a normal immune response. Antigens that can be altered to achieve this goal include, for example, MHC class I antigens, MHC class II antigens, LFA-3, and ICAM-1.

[0156] In another example, mesenchymal lineage precursor cells or stem cells can be genetically modified to express gene products that are supplied to the subject receiving the transplant. Examples of gene products that can be delivered to the subject by genetically modified mesenchymal lineage precursor cells include gene products that can prevent future cardiac disease, such as growth factors that promote blood vessels to invade the myocardium (e.g., vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF), transforming growth factor β (TGF-β) and angiotensin).

[0157] Mesenchymal lineage precursor cells or stem cells can also be genetically modified to express proteins important for differentiation and / or maintenance of striated skeletal muscle cells. Exemplary proteins include growth factors (TGF-β, insulin-like growth factor 1 (IGF-1), FGF), myogenic factors (e.g., myoD, myogenin, myogenic factor 5 (Myf5), myogenic regulatory factors (MRFs), transcription factors (e.g., GATA-4), cytokines (e.g., cardiotrophin-1), members of the neuregulin family (e.g., neuregulin 1, 2, and 3), and homeobox genes (e.g., Csx, tinman, and NKx families).

[0158] Heart failure

[0159] Heart failure occurs when the heart is unable to pump adequately to maintain blood flow to meet the body's needs. One cause of heart failure is myocardial infarction (MI). An MI occurs when blood stops flowing normally to a part of the heart. The lack of blood supply results in a localized area of ​​myocardial damage called an infarct or infarct formation. The infarcted heart is unable to pump adequately to maintain blood flow to meet the body's needs, resulting in heart failure. After an MI, a series of compensatory mechanisms are initiated to mitigate the drop in cardiac output and to help maintain adequate blood pressure to perfuse vital organs. As a result, patients with heart failure may not progress for a long time. However, the compensatory mechanisms are eventually unable to compensate for the damaged heart, resulting in a gradual drop in cardiac output, known as "progressive heart failure."

[0160] The diagnosis of MI is generated by integrating the history of presenting disease and physical examination with electrocardiographic findings and cardiac markers. Coronary angiography can be performed, which allows the visualization of stenosis or obstruction on the heart vessels. According to the WHO criteria revised in 2000 (Alpert JS, Thygesen K, Antman E, Bassand JP. (2000). "Myocardial infarction redefined--a consensus document of The Joint European Society of Cardiology / American College of Cardiology Committee for the redefinition of myocardial infarction". J Am Coll Cardiol 36 (3): 959–69), elevated cardiac troponin with typical symptoms, pathological Q waves, ST elevation or depression or coronary artery intervention is the diagnosis of MI.

[0161] For more than 70 years, the 12-lead electrocardiogram (ECG) remains the standard for determining the presence and location of MI. It is widely available, non-invasive, inexpensive and easily repeatable. The quantitative Selvester QRS scoring system (Selvester RH et al. (1985) Arch Intern Med 145 (10): 1877-1881) designed by computer simulation uses information on the ECG to estimate the size of MI. The Selvester scoring system is a 31-point QRS scoring system based on 50 criteria of observation of the Q- and R-wave durations and the R / Q and R / S amplitude ratios in the standard 12-lead electrocardiogram. Methods for determining infarct size, including but not limited to QRS scoring, are well known to those skilled in the art.

[0162] Cardiac markers may also be measured to determine the occurrence of MI. Such markers include troponin T and I, creatinine kinase, myoglobin levels, natriuretic peptides (e.g., B-type natriuretic peptide), C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), cardiac fatty acid binding protein and copeptin, intermediate regional preatrial natriuretic peptide, ST2, C-terminal proendothelin 1, and intermediate regional proadrenomedullin.

[0163] Troponin is a protein released from myocytes when irreversible myocardial damage occurs. It is highly specific for cardiac tissue and accurately diagnoses myocardial infarction in patients with a history of ischemic pain or ECG changes reflecting ischemia. Cardiac troponin levels are dependent on infarct size and therefore provide an indication of prognosis after infarction.

[0164] The methods of the present disclosure are directed to the treatment of the gradual decline in cardiac output characteristic of progressive heart failure.

[0165] Thus, in the context of this disclosure, "treat" and "treatment" refer to both therapeutic treatment and prophylactic or preventative measures.

[0166] In one example, treatment reduces the chance or risk of heart failure-related major adverse cardiac events (HF-MACE), which is defined as a composite of heart-related death or resuscitated cardiac death or non-fatal decompensated heart failure events. In one example, the risk chance of HF-MACE is reduced for at least 6 months, at least 12 months, at least 24 months, at least 36 months.

[0167] In the context of the present disclosure, it is contemplated that the terms chronic heart failure, congestive heart failure, and congestive heart failure may be used interchangeably with "progressive heart failure."

[0168] Myocardial infarction-induced heart failure

[0169] It is well known that patient progmosis and cardiac function are related to the amount of left ventricle (LV) infarcted.

[0170] The term "myocardial infarction-induced heart failure" refers to a subset of subjects in which myocardial infarction (MI) is the cause of heart failure. It is envisioned that the methods of the present disclosure can be used to treat progressive heart failure in a specific population of MI subjects.

[0171] In particular, the population of MI subjects is those with an LVESV greater than 70 mL. In other examples, 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 one example, the subject has a proximal left anterior descending (LAD) arterial lesion. In the context of the present disclosure, the term "arterial lesion" encompasses an obstructive lesion that obstructs the LAD of the heart; or an arterial lesion that previously obstructed the LAD that has been treated, for example, by percutaneous coronary intervention (PCI), also known as angioplasty.

[0172] MI can lead to persistent left ventricular dysfunction. Left ventricular dysfunction is characterized by a decrease in myocardial contractility. A decrease in left ventricular ejection fraction (LVEF) occurs when myocardial contractility decreases throughout the left ventricle. Therefore, LVEF provides a way to determine left ventricular dysfunction. LVEF can be measured by a variety of methods known in the art, such as, but not limited to, two-dimensional echocardiography (ECG), magnetic resonance tomography, cardiac computed tomography (CT), radionuclide angiography, gated myocardial perfusion single photon emission computed tomography (SPECT), gated myocardial perfusion positron emission tomography (PET), or biplane left ventricular cine angiography.

[0173] LVEF can be measured using the following formula: LVEF = stroke volume (EDV-ESV) / EDV. A change in LVEF of 5-10% may represent a true decrease in LVEF.

[0174] In one example, a subject having an LVEF of less than about 55% has left ventricular dysfunction. In other examples, a subject having an LVEF of less than about 54%, 53%, 52%, 51%, 50%, 49%, 48%, 47%, 46% has left ventricular dysfunction. In another example, a subject having an LVEF of less than about 45% has left ventricular dysfunction. In other examples, a subject having an LVEF of less than about 44%, 43%, 42%, 41% has left ventricular dysfunction. In another example, a subject having an LVEF of less than about 40% has left ventricular dysfunction.

[0175] In other examples, a subject having an LVEF less than about 39%, 38%, 37%, 36%, 35%, 34%, 33%, 32%, 31%, 30% has left ventricular dysfunction.

[0176] In the context of the present disclosure, the term "persistent left ventricular dysfunction" is used to define left ventricular dysfunction that persists over a period of time or a series of measurements. For example, "persistent left ventricular dysfunction" may include left ventricular dysfunction that persists between about 1 to about 14 days after MI, or longer. For example, persistent left ventricular dysfunction may include left ventricular dysfunction between about 1 to about 10, about 1 to about 9, about 2 to about 8, about 2 to about 7 days after MI. In another example, "persistent left ventricular dysfunction" may include left ventricular dysfunction that persists over about 1 to 10 or more measurements.

[0177] The size or amount of myocardial necrosis after MI is clinically referred to as infarct size. It is contemplated that the methods of the present disclosure involve treating MI subjects with large infarct sizes. For example, a subject treated using the methods of the present disclosure has an infarct size greater than about 10-35% of the left ventricle. In other instances, the subject has an infarct size greater than about 11-34%, about 12-33%, about 13-32%, about 14-31%, about 15-30%, about 16-29%, about 17-28% of the left ventricle. In another instance, the subject has an infarct size greater than about 18.5% of the left ventricle. In other instances, the subject has an infarct size of about 19-27%, about 20-26%, about 21-25%, about 22-24%, about 23% of the left ventricle.

[0178] Infarct size can be measured by a variety of methods known in the art. Examples of these methods include the use of serum markers such as creatine kinase (CK), CK-MB, troponin I, and brain natriuretic peptide troponin.

[0179] In one example, a subject treated with the methods of the present disclosure has a troponin level of at least about 2 x upper limit of normal (ULM).

[0180] In another example, the subject has a troponin level of at least about 3x, about 4x, about 5x, about 6x ULM.

[0181] In one example, a subject treated with the methods of the present disclosure has a creatine kinase-MB level of at least about 2 x upper limit of normal (ULM).

[0182] In another example, the subject has a creatine kinase-MB level of at least about 3x, about 4x, about 5x, about 6x ULM.

[0183] Other examples of measuring infarct size include Sestamibi single photon emission computed tomography (SPECT) myocardial perfusion imaging, magnetic resonance imaging. In one example, cMRI is used to measure infarct size. Several cardiac magnetic resonance imaging (cMRI) techniques can be used to diagnose infarct size. One of the most accurate and effective techniques is delayed enhancement cardiac magnetic resonance imaging (DE-CMR) technology. In one example, cMRI includes delayed enhancement cardiac magnetic resonance imaging (DE-CMR).

[0184] When using the appropriate settings of DE-CMR, normal myocardium appears black or ineffective, while non-vital areas appear bright or over-enhanced. Therefore, in one example, infarct size can be determined by visual assessment of bright and super-enhanced areas. Other examples of 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). In short, super-enhancement is scored on a 5-point scale for each segment on a 17-segment model (0 = no over-enhancement, 1 = 1% to 25%, 2 = 26% to 50%, 3 = 51% to 75%, 4 = 76% to 100%). The black area completely contained in the super-enhanced myocardium is interpreted as an area of ​​microvascular damage (no return) and is included in a portion of the infarct. Infarct size was calculated as a percentage of the LV myocardium by summing the regional scores, with each region weighted by the midpoint of the hyperenhancement range (i.e., 1 = 13%, 2 = 38%, 3 = 63%, 4 = 88%), and divided by 17. In another example, infarct size can be quantified by planimetry of the hyperenhanced area on the short-axis image stack.

[0185] In one example, infarct size is measured between about 1 day and 40 days after MI.

[0186] In other examples, infarct size is measured between about 1-40 days, between about 2-35 days, between about 3-30 days, between about 4-25 days, between about 5-20 days, between about 6-15 days, between about 6-15 days after MI.

[0187] In one example, infarct size can be measured about 30 days after MI.

[0188] 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 amount of the left ventricle that is infarcted.

[0189] It is contemplated that the methods of the present disclosure may be used to treat progressive heart failure in subjects with myocardial infarction having various stages or classifications of heart failure.

[0190] In one example, the heart failure staging is based on the American College of Cardiology (ACC) and American Heart Association (AHA) staging criteria. In another specific example, the subject has stage A, B, C or D heart failure according to the ACC or AHA criteria. In another example, the subject has stage B or C heart failure.

[0191] In another example, the heart failure classification is based on the New York Heart Association (NYHA) classification scale. In another specific example, the subject has class I, II, III or IV heart failure. In another example, the subject has stage II or III heart failure.

[0192] Catheter-based delivery systems

[0193] Any delivery system based on catheter can be used for implementing the method of the present disclosure, and the delivery system allows mesenchymal lineage precursor cell or stem cell and / or its offspring and / or soluble factors derived therefrom or the composition comprising it to be injected into the position near the myocardium of experimenter or cardiac tissue injury area. In some instances, catheter is introduced percutaneously (for example, enters femoral artery or another blood vessel), and is guided to the myocardium of experimenter by vascular system, in myocardium, it is used for delivering mesenchymal lineage precursor cell or stem cell and / or its offspring and / or soluble factors derived therefrom, or the composition comprising it via the needle extruded from catheter end. In other instances, catheter arrives at the heart by minimal surgical incision (for example, restrictive thoracotomy, it relates to the incision between ribs).

[0194] Several catheters have been designed to precisely deliver agents to damaged areas within the heart, such as infarcted areas (see, e.g., U.S. Pat. Nos. 6,102,926; 6,120,520; 6,251,104; 6,309,370; 6,432,119 and 6,485,481, the entire contents of each of which are incorporated herein by reference). The catheter can be guided to the specified location by downwardly conducting a steerable or guideable catheter having a receiving lumen (see, e.g., U.S. Pat. No. 5,030,204) or by a fixed structure guide catheter (see, e.g., U.S. Pat. No. 5,104,393). Alternatively, the catheter can be advanced to the desired location within the heart by a deflectable stylet (see, e.g., WO93 / 04724) or a deflectable guidewire (see, e.g., U.S. Pat. No. 5,060,660).

[0195] The catheter can be coupled to a cardiac positioning system that allows the location and extent of the damaged / defective area to be determined (as described above). Once the area in need of treatment has been identified, the steering guide can be pulled out, leaving the needle at the injection site. Some or all of the mesenchymal lineage precursor cells or stem cells and / or their progeny and / or their derived soluble factors are delivered down the lumen of the catheter and injected into the myocardium. The catheter is retracted from the subject when all injections have been performed.

[0196] When the catheter is guided into the heart of the subject, the needle element can be usually retracted in the sheath to avoid damaging the venous system and / or the myocardium. During injection, the needle is squeezed out from the tip of the catheter. During injection, the needle extends less than 10mm, less than 7.5mm or less than 5mm into the myocardial wall of an adult. Depending on the injection site, the maximum length can be changed. For infants and children, the extension depth is correspondingly smaller, as determined by the actual or estimated wall thickness. The needle specification for transplanting cells can be, for example, 25 to 30.

[0197] In one example, a catheter for delivering mesenchymal lineage precursor cells or stem cells and / or their offspring and / or soluble factors derived therefrom to the myocardium is configured to include a feedback sensor for locating the penetration depth and the position of the needle insertion. The use of a feedback sensor provides the advantage of accurately targeting the injection position. The target position for delivering the cell composition can vary. For example, optimal treatment may require multiple small injections in the injury / defect area without two injections penetrating the same part. Alternatively, the target position can remain the same in a continuous cell administration program.

[0198] A suitable catheter for use in the present disclosure is the NOGA TM Injection catheter system (Biosense Webster, Inc.). This catheter is a multi-electrode percutaneous catheter with a deflectable tip and an injection needle designed to inject medication into the myocardium. The tip of the injection catheter is equipped with a Biosense position sensor and a retractable hollow 27-gauge needle for fluid delivery. The injection site is indicated in real time on a cardiac map, allowing precise distribution of injections. Local electrical signals are obtained to minimize catheter tip trauma.

[0199] Compositions of the present disclosure

[0200] In one example of the present disclosure, mesenchymal lineage precursor cells or stem cells and / or their progeny and / or soluble factors derived therefrom are administered in the form of a composition. In one example, such a composition comprises a pharmaceutically acceptable carrier and / or excipient.

[0201] The terms "carrier" and "excipient" refer to compositions of matter that are routinely used in the art to facilitate the storage, administration, and / or biological activity of an active compound (see, e.g., Remington's Pharmaceutical Sciences, 16th edition, MacPublishing Company (1980)). The carrier may also reduce any undesirable side effects of the active compound. Suitable carriers are, for example, stable, e.g., unable to react with other components of the carrier. In one example, the carrier does not produce significant local or systemic adverse reactions in the recipient at the dose and concentration used for treatment.

[0202] Suitable carriers for the present disclosure include those conventionally used, for example, water, saline, aqueous dextrose, lactose, Ringer's solution, buffered solutions, hyaluronic acid and glycols are exemplary liquid carriers, particularly (when isotonic) for solutions. Suitable pharmaceutical carriers and excipients include starch, cellulose, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, magnesium stearate, sodium stearate, glyceryl monostearate, sodium chloride, glycerol, propylene glycol, water, ethanol, etc.

[0203] In another example, the carrier is a culture medium composition, such as a medium in which cells are cultured or suspended. For example, such a culture medium composition does not induce any adverse effects in a subject to which it is administered.

[0204] Exemplary carriers and excipients do not adversely affect the viability of the cells and / or the ability of the cells to reduce, prevent or delay metabolic syndrome and / or obesity.

[0205] In one example, the carrier or excipient provides buffering activity to maintain cells and / or soluble factors at a suitable pH to exert biological activity, such as a carrier or excipient is phosphate buffered saline (PBS). PBS represents an attractive carrier or excipient because it interacts minimally with cells and factors and allows rapid release of cells and factors, in which case the compositions of the present disclosure can be produced as a liquid for direct application to the blood, for example, by injection, or into tissues or areas surrounding or near tissues.

[0206] Mesenchymal lineage precursor cells or stem cells and / or their offspring and / or soluble factors derived therefrom may also be incorporated into or embedded into a scaffold that is compatible with a receptor and degrades into a product that is harmless to the recipient. These scaffolds provide support and protection for cells to be transplanted into a recipient subject. Natural and / or synthetic biodegradable scaffolds are examples of such scaffolds.

[0207] A variety of different scaffolds can be successfully used in the practice of the present disclosure. Exemplary scaffolds include, but are not limited to, biological, degradable scaffolds. Natural biodegradable scaffolds include collagen, fibronectin, and laminin scaffolds. Suitable synthetic materials for cell transplantation scaffolds should be able to support a wide range of cell growth and cell function. Such scaffolds can also be resorbable. Suitable scaffolds include polyglycolic acid scaffolds (e.g., as described in 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.

[0208] In another example, mesenchymal lineage precursor or stem cells and / or their progeny and / or soluble factors derived therefrom can be administered in a gel scaffold (such as Gelfoam from Upjohn Company).

[0209] The compositions described herein can be administered alone or as a mixture with other cells. Different types of cells can be mixed with the compositions of the present disclosure immediately or shortly before administration, or they can be co-cultured together for a period of time before administration.

[0210] In one example, the composition comprises an effective amount or a therapeutically or prophylactically effective amount of a mesenchymal lineage precursor cell or stem cell and / or its progeny and / or a soluble factor derived therefrom. For example, the composition comprises about 1x10 5 stem cells is about 1x10 9 stem cells or about 1.25x10 3 stem cells to about 1.25x10 7 The exact amount of cells to be administered depends on a variety of factors, including the age, weight, and sex of the subject and the extent and severity of the condition being treated.

[0211] Exemplary dosages include at least about 1.2 x 10 8 About 8x10 10 cells, such as about 1.3x10 8 About 8x10 9 cells, for example, about 1.4 x 10 8 About 8x10 8 cells, for example, about 1.5 x 10 8 About 7.2x10 8 cells, about 1.6x10 8 About 6.4x10 8 cells, such as about 1.7x108 to about 5.6x10 8 cells, for example, about 1.8 x 10 8 About 4.8x10 8 cells, for example, about 1.9 x 10 8 About 4x10 8 cells, about 2.0x10 8 About 3.2x10 8 cells, about 2.1x10 8 About 2.4x10 8 For example, a dose may include at least about 2.0 x 10 8 For example, a dose may include at least about 1.5 x 10 8 cells.

[0212] In other words, exemplary doses include at least about 1.5 x 10 6 cells / kg. For example, a dose may include about 1.5 x 10 6 About 1x10 9 cells / kg, such as about 1.6x10 6 About 1x10 8 cells / kg, such as about 1.8x10 6 About 1x10 7 cells / kg, for example, about 1.9 x 10 6 About 9x10 6 cells / kg, about 2.0x10 6 About 8x10 6 cells / kg, such as about 2.1x10 6 About 7x10 6 cells / kg, for example, about 2.3 x 10 6 About 6x10 6 cells / kg, for example, about 2.4 x 10 6 About 5x10 6 cells / kg, for example, about 2.5 x 10 6 About 4x10 6 cells / kg, for example, about 2.6 x 10 6 About 3x10 6 For example, a dose may include at least about 2.5 x 10 6 cells / kg.

[0213] In one example, mesenchymal lineage precursor cells or stem cells make up 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%, at least about 99% of the cell population of the composition.

[0214] The compositions of the present disclosure may be cryopreserved. Cryopreservation of mesenchymal lineage precursor cells or stem cells may be performed using slow cooling methods or "fast" freezing protocols known in the art. Preferably, the cryopreservation method maintains similar phenotypes, cell surface markers, and growth rates of cryopreserved cells compared to unfrozen cells.

[0215] The cryopreserved composition may comprise a cryopreservation solution. The pH of the cryopreservation solution is generally 6.5 to 8, preferably 7.4.

[0216] Intravenous injection solutions may include sterile, non-pyrogenic isotonic solutions such as PlasmaLyte A TM 100mL of PlasmaLyte A TM Contains 526 mg sodium chloride, USP (NaCl); 502 mg sodium gluconate (C6H 11 NaO7); 368 mg sodium acetate trihydrate, USP (C2H3NaO2·3H2O); 37 mg potassium chloride, USP (KCl); and 30 mg magnesium chloride, USP (MgCl2·6H2O). It does not contain an antimicrobial agent. The pH is adjusted with sodium hydroxide. The pH is 7.4 (6.5 to 8.0).

[0217] Cryopreservation solutions can include Profreeze TM The cryopreservation solution may additionally or alternatively comprise a culture medium, such as αMEM.

[0218] For the convenience of freezing, usually cryoprotectants, such as for example dimethyl sulfoxide (DMSO) are added to the cryopreservation solution. Ideally, cryoprotectants should be nontoxic, non-antigenic, chemically inert to cells and patients, provide high survival rate after thawing, and allow transplanting without cleaning. However, the most commonly used cryoprotectant DMSO shows some cytotoxicity. Hydroxyethyl starch (HES) can be used as a substitute or combined with DMSO to reduce the cytotoxicity of the cryopreservation solution.

[0219] The cryopreservation solution may include one or more of DMSO, hydroxyethyl starch, human serum components, and other protein fillers. In one example, the cryopreserved solution includes about 5% human serum albumin (HSA) and about 10% DMSO. The cryopreservation solution may also include one or more of methylcellulose, polyvinyl pyrrolidone (PVP), and trehalose.

[0220] In one embodiment, cells are suspended in 42.5% Profreeze TM / 50% αMEM / 7.5% DMSO and cooled in a controlled rate freezer.

[0221] The cryopreserved composition can be thawed and administered directly to a subject, or added to another solution (e.g., containing HA). Alternatively, the cryopreserved composition can be thawed and the mesenchymal lineage precursors or stem cells resuspended in an alternative carrier prior to administration.

[0222] In one example, a composition described herein can be administered from about 1 to about 10 days after MI.

[0223] In other examples, the compositions described herein can be administered about 1 to 9 days, about 1 to 8 days, about 2 to 7 days, about 2 to 6 days, about 3 to 5 days after MI. For example, the compositions described herein can be administered about 5 days after PCI.

[0224] In one example, the cell compositions described herein can be administered about 1 to about 10 days after percutaneous coronary intervention (PCI).

[0225] In other examples, the compositions described herein can be administered about 1 to 9 days, about 1 to 8 days, about 2 to 7 days, about 2 to 6 days, about 3 to 5 days after PCI. For example, the compositions described herein can be administered about 5 days after PCI.

[0226] In one example, a composition described herein can be administered in a single dose.

[0227] In some examples, the compositions described herein can be administered in multiple doses, for example, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10 doses.

[0228] In one example, mesenchymal lineage precursor cells or stem cells can be expanded in culture prior to administration. Various methods of cell culture are known in the art.

[0229] In one example, mesenchymal lineage precursor cells or stem cells are expanded by culturing in serum-free medium prior to administration.

[0230] In some instances, cells are contained within a chamber that does not allow cells to leave the subject's circulation, yet allows factors secreted by the cells to enter the circulation. In this manner, soluble factors can be administered to a subject by allowing cells to secrete factors into the subject's circulation. Such chambers can also be implanted at a site in a subject to increase local levels of soluble factors, such as in or near the heart.

[0231] Mesenchymal lineage precursor cells or stem cells and / or their progeny and / or soluble factors derived from the cells can be administered systemically, such as, for example, by intravenous, intraarterial or intraperitoneal administration. Mesenchymal lineage precursor cells or stem cells and / or their progeny and / or soluble factors derived therefrom can also be administered by intramuscular or intracardiac administration.

[0232] In one example, mesenchymal lineage precursor cells or stem cells and / or their progeny and / or soluble factors derived from the cells are administered directly into the myocardium. For example, mesenchymal lineage precursor cells or stem cells and / or their progeny and / or soluble factors derived from the cells can be administered directly into the myocardium of the left ventricle.

[0233] In one example, an intracardiac catheter such as the J&J Myostar TM The injection catheter administers mesenchymal lineage precursor or stem cells and / or their progeny and / or soluble factors derived from the cells.

[0234] In one example, mesenchymal lineage precursor or stem cells and / or their progeny and / or soluble factors derived from the cells are administered to viable myocardium.

[0235] In one example, mesenchymal lineage precursor or stem cells and / or their progeny and / or soluble factors derived from the cells are administered to hibernating myocardium.

[0236] Those skilled in the art will be able to identify viable and / or hibernating myocardium using methods known in the art. For example, a NOGASTAR TM Mapping catheter system Mapping catheter system to identify viable and / or hibernating myocardium.

[0237] In another example, mesenchymal lineage precursor cells or stem cells and / or their progeny and / or soluble factors derived from the cells are administered by intracoronary infusion. For example, mesenchymal lineage precursor cells or stem cells and / or their progeny and / or soluble factors derived from the cells can be administered to the left anterior descending artery (LAD) artery.

[0238] In one example, mesenchymal lineage precursor or stem cells and / or their progeny and / or soluble factors derived from the cells are administered into the LAD artery immediately following LAD revascularization by PCI.

[0239] 6-Minute Walk Test (6MWT)

[0240] The 6-minute walk test (6MWT) was developed in 1963 (Balke B et al. (1963) Rep Civ Aeromed Res Inst US 53:1-8) to assess exercise tolerance in chronic respiratory disease and heart failure. The test measures the distance a subject walks on a hard, flat surface in a total of 6 minutes. The goal is to walk as far as possible in six minutes. Subjects are allowed to pace on their own and rest as needed as they walk back and forth along a marked walkway.

[0241] Those skilled in the art will appreciate that many changes and / or modifications may be made to the above embodiments without departing from the broad overall scope of the present disclosure. Therefore, the present embodiments are considered in all aspects to be illustrative and not restrictive.

[0242] The following specific examples are to be construed as merely illustrative and not limiting in any way the remainder of the disclosure. Without further elaboration, it is believed that one skilled in the art can utilize the present invention to its fullest extent based on the description herein.

[0243] Example

[0244] Example 1: Study Design

[0245] A study was conducted to evaluate the safety and tolerability of 3 increasing doses (25, 75, or 150 million cells) of mesenchymal precursor cells (MPCs) in subjects with heart failure due to left ventricular systolic dysfunction of ischemic or nonischemic etiology. Secondary objectives were to observe efficacy by multiple parameters and to identify the optimal effective dose and optimal target population for MPC treatment.

[0246] Heart failure subjects were aged >18 years; heart failure due to ischemic or nonischemic cardiomyopathy, LV systolic dysfunction as demonstrated by ejection fraction (EF) <30% as measured on radionuclide ventriculography, two-dimensional echocardiography, or magnetic resonance imaging; and LV end-diastolic diameter >3.2 cm / m2 or >6 cm as determined by echocardiography; symptoms of dyspnea or fatigue at rest or with minimal exertion (New York Heart Association (NYHA) class II or III) for >2 months; at least one hospitalization or two outpatient visits requiring intravenous diuretic or vasodilator therapy within 12 months prior to screening; and optimal medical therapy, including diuretics, beta-blockers, and angiotensin-converting enzyme (ACE) inhibitors or angiotensin receptor blockers (ARBs), unless intolerable or contraindicated.

[0247] Subjects were excluded based on acute myocardial infarction within the previous 90 days, serum potassium <4.0 or >5.5 mEq / L, digoxin level >1.2 ng / mL, magnesium level <1.0 mEq / L, serum creatinine >2.0 mg / dL, and serum bilirubin >3.0 mg / dL.

[0248] Subjects were randomized to receive an injection of 25 million, 75 million, or 150 million MPCs based on either an endomyocardial catheter or a written sham injection (control group) in the catheterization laboratory. TM Injection catheter and NOGASTAR TM The mapping catheter system applies MPC to the left ventricle (about 15-20 injections of 0.2ml / injection), and the system identifies vitality / hibernation myocardium based on voltage, theoretically making the targeting of healthy but risky tissues easier. The catheter has the maximum safety profile of this application, and has been used in more than 1,000 patients in multiple trials. The measurement of functional effectiveness involves left ventricular end-systolic volume or LVESV, left ventricular end-diastolic volume or LVEDV, measurement and left ventricular ejection fraction or LVEF. The other analysis of the time before the first event of the major adverse cardiac events related to heart failure or HF-MACE was carried out. HF-MACE is defined as the composite of heart-related death or resuscitation cardiac death or non-fatal decompensated heart failure events.

[0249] Conclusions: Endomyocardial injection of MPC in patients with chronic heart failure is feasible and safe. The incidence of adverse events was similar in all groups, and there were no clinically significant immune responses in any patient receiving MPC.

[0250] The 150 million cell dose showed the greatest effect on left ventricular remodeling and functional capacity and a threshold benefit for reducing long-term HF-MACE ( Figure 1 ).

[0251] More specifically, there was a dose-related effect between both LVESV and LVEDV, with the 150 million cells dose limiting the greatest effect compared to control for LV remodeling at 6 months after treatment (p<0.02 for both LVESV and LVEDV) and functional exercise capacity measured by the 6-minute walk test at 12 months after treatment (6MTW: p=0.062). The p value is a probability, ranging from 0 to 1, which indicates the likelihood that the results of the study are different between the treatment and control groups. The lower the p value, the harder it is to see the results by chance alone. P values ​​below 0.05 are generally referred to as statistically significant.

[0252] Independent blinded adjudication of potential HF-MACE was performed post hoc. The 150 million cell dose was associated with a significantly greater likelihood of remaining free of HF-MACE events compared to the control group over the 36-month follow-up (0% vs. 33% HF-MACE by Kaplan-Meier, p=0.026 by log-rank test). The 25 and 75 million doses were not statistically different from the control group on this measure. On the basis of these results, the optimal dose for therapeutic benefit was considered to be the 150 million MPC dose ( Figure 2 ).

[0253] Example 2: Therapeutic Benefit of MPC Administration on Parameters of Cardiac Function (Intention to Treat Analysis)

[0254] In order to identify the most appropriate target population for the 150 million MPC dose, the inventors evaluated whether the best responders to MPC therapy were in groups with more or less advanced heart failure. Further post hoc analyses were performed in a blinded manner, stratifying patients in the control group or 150 million MPC treatment into patients with a baseline LVESV of less than or greater than 100 ml as an alternative to significant myocardial contractile abnormalities and advanced heart failure. The 100 ml LVESV threshold was selected because it fell within more than 3 standard deviations above the normal LVESV.

[0255] Table 1 below shows an analysis of the comparison of all subjects (i.e., subjects with any value of LVEF) to subjects with LVESV>100 in the intended treatment population. Intent to treat analysis is a statistical concept. According to Fisher et al. (Fisher LD, Dixon DO, Herson J, Frankowski RK, Hearron MS, Peace KE. Intention to treat in clinical trials. In: Peace KE ed. Statistical issues in drug research and development. New York: Marcel Dekker; 1990. Pages 331–50 (1990)), the ITT analysis includes all randomized patients in the group to which they are randomly assigned, regardless of whether they comply with the inclusion criteria, regardless of the treatment they actually receive, and regardless of whether they subsequently withdraw from treatment or deviate from the protocol. In other words, the ITT analysis includes each subject randomized according to the randomized treatment assignment. It ignores noncompliance, protocol deviation, withdrawal, and anything that occurs after randomization. ITT analysis avoids overly optimistic estimates of the efficacy of an intervention resulting from the removal of non-compliers by accepting that noncompliance and protocol deviations may occur in actual clinical practice.

[0256] Table 1 Comparison of all subjects with subjects with LVESV>100 ml (ITT, or intent to treat)

[0257]

[0258] This analysis demonstrated that the treatment benefit of the 15,000 dose on parameters of LV remodeling was significantly amplified by focusing on a target population with substantial baseline LV systolic abnormalities and advanced heart failure (LVESV greater than 100 ml).

[0259] Example 3: Therapeutic Benefit of MPC Administration on LVESV

[0260] A total of 30 subjects (mixed with heart failure and non-heart failure subjects) were assessed for baseline LVESV levels. The distribution of subjects is shown in Table 2. Subjects were classified into placebo group or MPC cell group (which was administered 1.5x10 8 Mesenchymal progenitor cells (MPCs) were injected into the left ventricle (approximately 15-20 injections of 0.2 ml / injection) using the J&J Myostar injection catheter and the NOGASTAR mapping catheter system for voltage-based identification of viable / hibernating myocardium. Subjects were stratified based on their left ventricular end-systolic volume (LVESV) values ​​≤100 mL or >100 mL.

[0261] Table 2 Stratification of subjects according to baseline LVESV value

[0262]

[0263] Of the 30 subjects evaluated for the study, 18 subjects had baseline LVESV values ​​> 100 mL. Seven (7) subjects were assigned to placebo and 11 were assigned to MPC.

[0264] Figure 3-1 Baseline LVESV values ​​for the subject groups are shown. Subjects stratified according to LVESV less than or equal to (≤) 100 mL had a mean baseline LVESV value of 81 mL. Subjects stratified according to their LVESV values ​​greater than (>) 100 mL had a mean baseline value of 136 mL.

[0265] After administration of placebo or 1.5x10 8 Subjects were re-evaluated at the 6-month time point after the initiation of mesenchymal progenitor cell (MPC) injection. Major adverse cardiac event (MACE) rates were determined for subjects stratified based on LVESV values. Figure 3-2 Results are shown in , representing the change in LVESV values ​​between baseline and 6-month time points. The change in LVESV values ​​between subjects administered placebo (control) and MPC was statistically significant.

[0266] Table 3 shows the heart failure MACE rates (HF-MACE) expressed as numbers and percentages.

[0267] Table 3 Changes in LVESV values ​​among subjects stratified according to baseline LVESV values

[0268]

[0269]

[0270] Subjects with baseline LVESV values ​​≤100 mL had no major adverse cardiac events during the 6-month evaluation period, which was independent of whether the subjects were treated with placebo or 1.5x10 8 HF-MACE occurred only in placebo subjects (controls) with evidence of greater baseline cardiac dysfunction, as assessed by LVESV>100 mL. In contrast, none of the subjects with baseline cardiac dysfunction, as assessed by LVESV>100 mL, who were administered MPCs developed HF-MACE.

[0271] The data in Table 2 indicate that heart failure subjects treated with placebo will experience high MACE rates (ie, 71% over a 6-month period). In particular, high MACE rates were only observed in subjects with baseline LVESV values ​​>100 mL.

[0272] The data showed that heart failure subjects with baseline LVESV values ​​>100 mL derived the best therapeutic benefit from the administration of MPC. In other words, subjects at the highest risk of disease progression derived the greatest benefit from the administration of MPC.

[0273] Example 4: Therapeutic Benefits of MPC Administration on LVEDV and LVEF

[0274] Left ventricular end-diastolic volume (LVEDV) was also assessed in subjects who had been stratified according to LVESV values ​​less than or equal to (≤) 100 mL or >100 ml.

[0275] Figure 4-1 The subjects' baseline LVEDV values ​​are shown. The mean baseline LVEDV for subjects with a baseline LVESV ≤ 100 mL was 128 mL. The mean baseline LVEDV for subjects with a baseline LVESV > 100 mL was 198 mL.

[0276] The distribution of subjects is as shown in Table 1.

[0277] Figure 4-2 Shown is the change of LVEDV value after 6 months of placebo (control) or MPC administration between the subjects stratified according to baseline LVESV≤100mL and baseline LVESV>100mL.This figure shows that the heart failure subjects with the highest MACE rate achieve the therapeutic benefit from MPC administration, as evidenced by the improvement of LVEDV value.The difference between the subjects stratified according to LVESV value≤100mL or>100mL is statistically significant.

[0278] Left ventricular ejection fraction (LVEF) values ​​were also assessed in subjects stratified by LVESV values. Figure 5-1 The subjects' baseline LVEF values ​​are shown. Subjects with baseline LVESV ≤ 100 mL had a mean baseline LVEF of 35-40%. Subjects with baseline LVESV values ​​> 100 mL had a mean baseline LVEF value of approximately 32%.

[0279] The distribution of subjects is as shown in Table 1.

[0280] Figure 5-2Shown is the change of LVEF value after 6 months of applying placebo (control) or MPC between the subjects stratified according to baseline LVESV≤100mL and baseline LVESV>100mL.This figure shows that the heart failure subjects with the highest MACE rate reach the therapeutic benefit from MPC administration, as demonstrated by the improvement of LVEF value.The difference between the subjects stratified according to LVESV value≤100mL or>100mL is statistically significant.

[0281] Example 5: Correlation between disease severity and therapeutic benefit of MPC for LVESV

[0282] Further sensitivity analyses across each decile of baseline LVESV between 70 ml and 120 ml confirmed the findings seen in the stratification using LVESV greater than 100 ml.

[0283] Figures 6 to 11 The results showed that the effects of placebo (control) or MPC (1.5x10 8 The change in LVESV of subjects evaluated 6 months after the induction of 100% mesenchymal progenitor cells (100% mesenchymal progenitor cells) was measured. The reduction in LVESV correlated with the level of heart failure (as determined by the measurement of baseline LVESV). Subjects were stratified according to LVESV as shown in Table 4 below.

[0284] Table 4 Changes of LVESV

[0285]

[0286] Example 6: HF-MACE in patients with LVESV>100 ml.

[0287] HF-MACE was examined 36 months after treatment in the control group and in subjects who received 150 million MPCs. All HF-MACE events within 36 months of follow-up occurred only in the control group with advanced heart failure ( Fig.12 ).

[0288] The annualized HF-MACE rate among these rapid progressors was 24%, compared with 11% among all controls.

[0289] More specifically, among 18 Class II / III CHF patients with baseline LVESV>100 ml, 5 / 7 (71%) placebo-treated vs. 0 / 11 150 MPC-treated patients experienced one or more HF-MACE events within 36 months (p=0.0007).

[0290] Therefore, the effect of the 150 million MPC dose on overall HF-MACE was significantly amplified in those patients with advanced heart failure and high progression rate, and this may represent the optimal target patient population for MPC therapy.

[0291] Comments

[0292] The data shown herein show that the greater the magnitude of baseline left ventricular contractile abnormalities in chronic heart failure subjects due to left ventricular systolic dysfunction, the more beneficial the MPC-related cardioprotective effects observed during the 6-month follow-up period. The data further show that the progressive adverse natural history associated with advanced chronic heart failure can be beneficially changed by treatment with MPC. Without wishing to be bound by theory, it is found that the paracrine crosstalk hypothesis is supported, in which biochemical / physiological disturbances at the tissue level produce a local environment that promotes MPC to release beneficial paracrine mediators. Therefore, the best effect obtained by administering MPC in heart failure subjects is observed in subjects with the highest risk of disease progression, i.e., subjects with baseline LVESV>70mL.

[0293] Baseline LVESV>70 ml identified a rapidly progressive subgroup of subjects with chronic heart failure and left ventricular systolic dysfunction who experienced high HF-MACE.

[0294] In subjects with baseline LVESV >70 ml, >80 ml, >90 ml, >100 ml, >110 ml, or >120 ml, treatment with high-dose (150 million) MPCs resulted in greater improvements in cardiac remodeling variables and HF-MACE than seen in control subjects.

[0295] The results presented herein identify the optimal target population for the cardioprotective benefit of cell therapy and facilitate improvements in trial designs for subjects with heart failure due to left ventricular systolic dysfunction.

[0296] Control patients with advanced heart failure (baseline LVESV>100 ml) were the most rapid progressors over 6 months with significant deterioration in LVESV and LVEDV volumes and loss of LVEF. During the 6-month follow-up period, the 150 million MPC dose had a significant cardioprotective effect on LVESV (p<0.02), LVEDV (p<0.03), and LVEF (p<0.05) in class II / III patients with substantial baseline LV contractile abnormalities (i.e., those with baseline LVESV>100 ml).

[0297] During the 6-month follow-up period, the 150 million MPC dose had significant cardioprotective effects on LVESV (p<0.02), LVEDV (p<0.03), and LVEF (p<0.05) in class II / III patients with substantial baseline LV systolic abnormalities (i.e., those with baseline LVESV>100 ml).

[0298] This application relates to the following implementation schemes:

[0299] 1. A method for treating or preventing heart failure in a subject in need thereof, the method comprising administering to the subject a population of mesenchymal lineage precursor or stem cells and / or progeny thereof and / or soluble factors derived therefrom, the subject having an elevated left ventricular end-systolic volume (LVESV) greater than 70 mL.

[0300] 2. The method of embodiment 1, wherein the elevated LVESV is due to acute myocardial infarction.

[0301] 3. The method of embodiment 1, wherein the elevated LVESV is due to chronic congestive heart failure.

[0302] 4. The method according to any one of embodiments 1 to 3, comprising the following steps:

[0303] i) select subjects with an LVESV greater than 70 mL, and

[0304] ii) administering to the subject a population of mesenchymal lineage precursor or stem cells and / or progeny thereof and / or soluble factors derived therefrom.

[0305] 5. The method according to any preceding embodiment, wherein the subject has a LVESV greater than 80 mL.

[0306] 6. The method according to any preceding embodiment, wherein the subject has a LVESV greater than 90 mL.

[0307] 7. The method according to any preceding embodiment, wherein the subject has a LVESV greater than 100 mL.

[0308] 8. The method according to any preceding embodiment, wherein the subject has a LVESV greater than 110 mL.

[0309] 9. The method according to any preceding embodiment, wherein the cells and / or their progeny and / or soluble factors are administered to the subject via a catheter-based system.

[0310] 10. The method of embodiment 9, wherein the cells and / or their progeny and / or soluble factors are administered to the subject by transendocardial injection, intracoronary infusion, intravenous infusion, or transepicardial injection.

[0311] 11. The method according to any preceding embodiment, wherein the subject is characterized as having a left ventricular ejection fraction (LVEF) of less than or equal to about 35%.

[0312] 12. The method of embodiment 1, wherein the heart failure is due to hypertension, cardiomyopathy (ischemic or non-ischemic), myocarditis, obesity or diabetes.

[0313] 13. The method according to any preceding embodiment, wherein the population of mesenchymal lineage precursor or stem cells and / or progeny thereof and / or soluble factors derived therefrom are administered to the subject about 1-7 days after diagnosis of heart failure.

[0314] 14. The method according to any preceding embodiment, wherein the subject has an infarct size of approximately 5%-30% of the left ventricle.

[0315] 15. A method for treating or preventing heart failure in a heart having an LVESV greater than 70 mL, the method comprising administering to the heart a population of mesenchymal lineage precursor or stem cells and / or progeny thereof and / or soluble factors derived therefrom

[0316] 16. The method of embodiment 15, wherein the heart has a LVESV greater than 100 mL.

[0317] 17. The method according to any preceding embodiment, comprising administering a population of mesenchymal lineage precursor or stem cells enriched for STRO-1+ cells and / or their progeny and / or soluble factors derived therefrom.

[0318] 18. The method according to any of the preceding embodiments, comprising administering a monoclonal antibody directed against STRO-1. 亮 Cells are enriched for populations of mesenchymal lineage precursor or stem cells and / or their progeny and / or soluble factors derived therefrom.

[0319] 19. The method according to any preceding embodiment, wherein the population of mesenchymal lineage precursor or stem cells and / or progeny thereof are administered to the subject or the heart in multiple doses.

[0320] 20. The method according to any preceding embodiment, comprising administering about 1.5 x 10 8 cells and / or their progeny.

[0321] 21. The method according to any preceding embodiment, wherein the population of mesenchymal lineage precursor or stem cells and / or their progeny are administered as a composition together with a pharmaceutically acceptable carrier and / or excipient.

[0322] 22. The method according to any preceding embodiment, wherein the population of mesenchymal lineage precursor or stem cells and / or their progeny have been culture expanded prior to administration and / or prior to obtaining the soluble factors.

[0323] 23. Use of a population of mesenchymal lineage precursor or stem cells and / or progeny thereof and / or soluble factors derived therefrom, or compositions thereof, for treating heart failure in a subject with an elevated LVESV greater than 70 mL.

[0324] 24. Use of a population of mesenchymal lineage precursor or stem cells and / or progeny thereof and / or soluble factors derived therefrom, or compositions thereof, for treating a heart having an elevated LVESV greater than 70 mL.

[0325] 25. The use according to embodiment 23 or 24, wherein the elevated LVESV is due to acute myocardial infarction.

[0326] 26. The use according to embodiment 23 or 24, wherein the elevated LVESV is due to chronic congestive heart failure.

[0327] 27. The method or use according to any preceding embodiment, wherein the mesenchymal lineage precursor or stem cells and / or their progeny are autologous or allogeneic.

[0328] 28. The method or use according to any preceding embodiment, wherein the subject is classified as New York Heart Association (NYHA) class II, III or IV.

[0329] 29. The method or use according to any preceding embodiment, wherein conventional heart failure medications are also concurrently administered to the subject.

[0330] 30. A population of mesenchymal lineage precursor or stem cells and / or progeny thereof and / or soluble factors derived therefrom for use in the manufacture of a medicament for treating heart failure in a subject having a left ventricular end-systolic volume (LVESV) greater than 70 mL.

[0331] 31. A population of mesenchymal lineage precursor or stem cells and / or progeny thereof and / or soluble factors derived therefrom for use in the manufacture of a medicament for treating a heart having a left ventricular end-systolic volume (LVESV) greater than 70 mL.

[0332] 32. A kit comprising a population of mesenchymal lineage precursor or stem cells and / or progeny thereof and / or soluble factors derived therefrom or a composition thereof, and a delivery device for administering the cells, and / or progeny thereof and / or soluble factors.

Claims

1. Enriched STRO-1+ cells 1.3 × 10 8 Up to 8×10 10 Use of a population of culture-expanded mesenchymal lineage precursor cells (MPCs) in the preparation of a medicament for treating or preventing heart failure caused by left ventricular systolic dysfunction in a subject in need thereof, wherein the subject is characterized by: (i) have an elevated baseline left ventricular end-systolic volume (LVESV) greater than 100 mL, and (ii) Baseline left ventricular ejection fraction (LVEF) less than or equal to 35%.

2. The use according to claim 1, wherein the LVESV of the subject measured at 6 months after administration of the MPC is reduced by at least 8 ml relative to the subject's baseline LVESV.

3. The use according to claim 1, wherein the left ventricular end-diastolic volume (LVEDV) of the subject measured at 6 months after administration of the MPC is reduced by at least 10 ml relative to the subject's baseline LVEDV.

4. The use according to claim 1, wherein the population comprises 1.3×10 8 to 8×10 9 MPC cells.

5. The use according to claim 1, wherein the elevated LVESV is due to chronic congestive heart failure.

6. The use according to claim 1, wherein the subject has a LVESV greater than 110 mL.

7. The use according to claim 1, wherein the cells are administered to the subject via a catheter-based system.

8. The use according to claim 7, wherein the cells are administered to the subject by transendocardial injection, intracoronary infusion, intravenous infusion or transepicardial injection.

9. The use according to claim 1, wherein the heart failure is due to hypertension, ischemic or non-ischemic cardiomyopathy, myocarditis, obesity or diabetes.

10. The use according to claim 1, wherein the mesenchymal lineage precursor cells are administered to the subject 1-7 days after diagnosis of heart failure.

11. The use according to claim 1, wherein the subject has an infarct size of 5%-30% of the left ventricle.

12. The use according to claim 1, wherein the cells are enriched for STRO-1 亮 cell.

13. The use according to claim 1, wherein the population comprises 1.5×10 8 to 8×10 9 MPCs.

14. The use according to claim 1, wherein the population of culture-expanded mesenchymal lineage precursor cells is administered in the form of a composition together with a pharmaceutically acceptable carrier and / or excipient.

15. The use according to claim 1, wherein the medicament improves LVESV and left ventricular end-diastolic volume (LVEDV) of the subject at 6 months after administration of the MPC.

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