Enhancing Myocardial Repair with CardioClusters

Abstract

1Background: Cellular therapy to treat heart failure is an ongoing focus of intense 2 research and development, but progress has been frustratingly slow due to limitations of 3 current approaches. Engineered augmentation of established cellular effectors 4 overcomes impediments, enhancing reparative activity with improved outcomes relative 5 to conventional techniques. Such 'next generation' implementation includes delivery of 6 combinatorial cell populations exerting synergistic effects. Concurrent isolation and 7 expansion of three distinct cardiac-derived interstitial cell types from human heart tissue, 8 as previously reported by our group, prompted design of a three-dimensional (3D) 9 structure that maximizes cellular interaction, allows for defined cell ratios, controls size, 10 enables injectability, and minimizes cell losses upon delivery. 11 Methods: Three distinct populations of human cardiac interstitial cells including 12 mesenchymal stem cells (MSCs), endothelial progenitor cells (EPCs), and c-Kit + cardiac 13 interstitial cells (cCICs) when cultured together spontaneously form scaffold-free 3D 14 microenvironments termed CardioClusters. Biological consequences of CardioCluster 15 formation were assessed by multiple assays including single cells RNA-Seq 16 transcriptional profiling. Protective effects of CardioClusters in vitro were measured 17 using cell culture models for oxidative stress and myocardial ischemia in combination 18 with freshly isolated neonatal rat ventricular myocytes. Long-term impact of adoptively 19 transferred CardioClusters upon myocardial structure and function in a xenogenic model 20 of acute infarction using NOD scid mice was assessed over a longitudinal time course of 21 20-weeks. 22 Results: CardioCluster design enables control over composite cell types, cell ratios, 1 size, and preservation of structural integrity during delivery. Profound changes for 2 biological properties of CardioClusters relative to constituent parental cell populations 3 include enhanced expression of stem cell-relevant factors, adhesion/extracellular-matrix 4 molecules, and cytokines. The CardioCluster 3D microenvironment maximizes cellular 5 interaction while maintaining a more native transcriptome similar to endogenous cardiac 6 cells. CardioCluster delivery improves cell retention following intramyocardial injection 7 with preservation of long-term cardiac function relative to monolayer-cultured cells when 8 tested in an experimental murine infarction model followed for up to 20 weeks post-9 challenge. CardioCluster-treated hearts show increases in capillary density, 10 preservation of cardiomyocyte size, and reduced scar size indicative of blunting 11 pathologic infarction injury. 12 Conclusions: CardioClusters are a novel 'next generation' development and delivery 13 approach for cellular therapeutics that potentiate beneficial activity and enhance 14 protective effects of human cardiac interstitial cell mixed populations. CardioClusters 15 utilization in this preclinical setting establishes ...

Hybrid SMART spheroids to enhance stem cell therapy for CNS injuries

“…Stem cell spheroids, which are 3D spherical cellular assemblies, have recently garnered much attention as promising avenues for disease modeling, drug discovery, and stem cell therapy (34)(35)(36). Typically generated from 3D cell culture, spheroid-based scaffold-free stem cell therapies can enhance cell survival and differentiation and stimulate the secretion of neurotrophic factors through their biomimetic 3D cell-cell interactions (30,31,37,38). They also allow the injectable delivery of high densities of stem cells at sites of CNS injuries or diseases in an accurate and relatively noninvasive manner, thereby enhancing the therapeutic outcomes of stem cell implantation (29,30,32,33).…”

hESCs‐Derived Early Vascular Cell Spheroids for Cardiac Tissue Vascular Engineering and Myocardial Infarction Treatment

“…Apart from engineering pre‐vascularized cardiac engraftment, cell therapy strategy with cell sheets or cell aggregates rather than singularized cells can also boost cell engraft rate and survival rate in injured heart. [ 13,38 ] Preclinical study has shown that MI therapy with hESCs‐derived CMs in non‐human primates has a very low survival rate (7%), which is in part due to the high pressure and continuous contraction of the pumping heart. [ 31 ] Alternatively, cell delivery with cell sheets or cell aggregates can be more effective.…”

“…[ 31 ] Alternatively, cell delivery with cell sheets or cell aggregates can be more effective. [ 13,38 ] We assumed that EVC spheroids may also be superior to EVC single cells in the treatment of myocardial infarction. Our results indicated that to a certain extent, EVC spheroids may be more helpful for cardiac function recovery after injection into infarcted area, which suggested that cell aggregates can be an alternative cell delivery strategy for cell therapy.…”

A Topologically Engineered Gold Island for Programmed In Vivo Stem Cell Manipulation

“…They mainly rely on the cleavage or concealing of the RGD ligands in response to magnetism, [23–25] electrochemical potential, [26] light, [27–31] pH, [32, 33] and enzymes [34, 35] . However, the stem cells harvested by these methods lack the capacity to sense and adhere to the target tissues, therefore requiring further complex modification in vitro to introduce targeted components, such as magnetic nanoparticles (MNPs) [36–41] . The post‐collection modification of cells undoubtedly increases the difficulty of cell manipulation and medical costs.…”