Transplantation of endothelial progenitor cells (EPCs) is a proven safe and effective method for treatment of cerebral ischemia in animal experiments. However, safety and efficacy need to be determined in clinical trials. We performed a two‐center, randomized, placebo‐controlled phase I/IIa trial with blinded outcome assessment on 18 patients with acute cerebral infarct within the middle cerebral artery territory, and followed for up to 4 years. Autologous ex vivo expanded EPCs were injected intravenously in the EPC group, and patients who received saline or autologous bone marrow stromal cells served as control groups. Mortality of any cause, adverse events, and new‐onset comorbidities were monitored. Changes in neurological deficits were assessed at different time points. We found no toxicity events or infusional or allergic reactions in any treated group. Three patients in the placebo group died during the 4‐year follow‐up. We found that the EPC group had fewer serious adverse events compared with the placebo‐controlled group, although there were no statistical differences in mortality among the three groups. Furthermore, there was no significant difference in neurological or functional improvement observed among the three groups, except for the Scandinavia Stroke Scale score at 3 months between the EPC group and placebo‐controlled group. Autologous transplantation of EPCs appears to improve long‐term safety in acute cerebral infarct patients, supporting the feasibility of this novel method for treatment of ischemic stroke. Stem Cells Translational Medicine 2018

The human isocitrate dehydrogenase (IDH) gene encodes for the isoenzymes IDH1, 2, and 3, which catalyze the conversion of isocitrate and α-ketoglutarate (α-KG) and are required for normal mammalian metabolism. Isocitrate dehydrogenase 1 and 2 catalyze the reversible conversion of isocitrate to α-KG. Isocitrate dehydrogenase 3 is the key enzyme that mediates the production of α-KG from isocitrate in the tricarboxylic acid (TCA) cycle. In the TCA cycle, the decarboxylation reaction catalyzed by isocitrate dehydrogenase mediates the conversion of isocitrate to α-KG accompanied by dehydrogenation, a process commonly known as oxidative decarboxylation. The formation of 6-C isocitrate from α-KG and CO₂ catalyzed by IDH is termed reductive carboxylation. This IDH-mediated reversible reaction is of great importance in tumor cells. We outline the role of the various isocitrate dehydrogenase isoforms in cancer, discuss the metabolic implications of interference with IDH, summarize therapeutic interventions targeting changes in IDH expression, and highlight areas for future research.

Acute ischemic stroke (AIS) is caused by clotting in the cerebral arteries, leading to brain oxygen deprivation and cerebral infarction. Recombinant human tissue plasminogen activator (tPA) is currently the only FDA-approved drug for ischemic stroke. However, tPA has to be administered within 4.5 h from the disease onset and delayed treatment of tPA can increase the risk of neurovascular impairment, including neuronal cell death, blood-brain barrier (BBB) disruption, and hemorrhagic transformation. A key contributing factor for tPA-induced neurovascular impairment is activation of matrix metalloproteinase-9 (MMP-9). We used a clinically-relevant mouse embolic model of focal-cerebral ischemia by insertion of a single embolus of blood clot to block the right middle cerebral artery. We showed that administration of the potent and highly selective gelatinase inhibitor SB-3CT extends the time window for administration of tPA, attenuating infarct volume, mitigating BBB disruption, and antagonizing the increase in cerebral hemorrhage induced by tPA treatment. We demonstrated that SB-3CT attenuates tPA-induced expression of vascular MMP-9, prevents gelatinase-mediated cleavage of extracellular laminin, rescues endothelial cells, and reduces caveolae-mediated transcytosis of endothelial cells. These results suggest that abrogation of MMP-9 activity mitigates the detrimental effects of tPA treatment, thus the combination treatment holds great promise for extending the therapeutic window for tPA thrombolysis, which opens the opportunity for clinical recourse to a greater number of patients. Significance Statement Endovascular thrombectomy benefited patients with disproportionately severe clinical deficit relative to infarct volume and provides a surgical option for treatment of ischemic stroke. Tissue-plasminogen activator (tPA) is the only FDA-approved clot-dissolving drug for treatment of ischemic stroke. However, its beneficial effect is seen only within 4.5 hours from the onset of stroke, with better outcomes observed the sooner treatment is given. Major risk is MMP-9−mediated neurovascular impairment. We document that early administration of the pharmacological agent SB₃CT reduces MMP-9 levels and mitigates the side effects associated with tPA. Furthermore, it extends the time window for treatment with tPA after stroke holding great potential for both disease intervention and benefiting patients who might not otherwise be candidates for treatment with tPA.

Expansion of bone marrow-derived endothelial progenitor cells (EPCs) in vitro to obtain required cell numbers for therapeutic applications faces the challenge of growing cell senescence under the traditional normoxic culture condition. We previously found that 1% O₂ hypoxic culture condition is favorable for reducing senescence of EPCs, but the mechanisms underlying the favorability are still unclear. Here, we found that, compared with normoxia, hypoxia induced a shift in lactate dehydrogenase (LDH) isozyme profile, which manifested as decreased LDH2 and LDH1 and increased LDH5, LDH4 and total LDHs. Moreover, under hypoxia, EPCs presented higher LDH activity, which could promote the conversion of pyruvate to lactate, as well as a higher level of NAD⁺, Bcl2 interacting protein 3 (BNIP3) expression and mitophagy. Additionally, under hypoxia, knock-down of the LDHA subunit increased the LDH2 and LDH1 levels and knock-down of the LDHB subunit increased the LDH5 level, while the simultaneous knock-down of LDHA and LDHB reduced total LDHs and NAD⁺ level. Inhibition of NAD⁺ recycling reduced BNIP3 expression and mitophagy and promoted cell senescence. Taken together, these data demonstrated that 1% O₂ hypoxia induces a shift in the LDH isozyme profile, promotes NAD⁺ recycling, increases BNIP3 expression and mitophagy, and reduces EPC senescence. Our findings contribute to a better understanding of the connection between hypoxic culture conditions and the senescence of bone marrow-derived EPCs and provide a novel strategy to improve in vitro expansion of EPCs.

Recent studies have suggested that bone marrow stromal cells (BMSC) have the potential to differentiate into endothelial cells. However, the physiologic functions of the endothelial-like cells derived from BMSC have not been well studied. Human BMSC were induced to differentiate into endothelial-like cells with a combination of cytokines. Morphologic, phenotypic, ultrastructural and functional characterizations of the endothelial-like cells were made. Human BMSC were successfully differentiated into cells with endothelial-like morphology and phenotype in vitro. These cells expressed various endothelial cell functions in vitro, such as release of von Willebrand factor (vWF) mediated by histamine, acetylated low-density lipoprotein (acLDL) uptake, binding of Ulex europaeus agglutinin-1 (UEA-1) and in vitro capillary formation. The cells also acquired important ultrastructural and physiologic properties of endothelial cells as they contained Weibel–Palade bodies, abundant mitochondria with a homogeneous mitochondrial matrix, diluted rough endoplasmic reticula, enlarged Golgi complexes, a regular arrangement of microfilaments and many surface cytoplasmic processes and plasmalemmal vesicles, as well as intercellular tight junctions and desmosome-like structures. Subcutaneous implantation of the endothelial-like cells in Matrigel plugs in immunodeficient mice resulted in the formation of functional blood vessels that contained erythrocytes. Moreover, these cells contributed to in vivo neovascularization during wound healing in rabbit ischemic hindlimb models. Physiologic features of the endothelial-like cells derived from BMSC suggest the potential use of these cells as a functional cell source for therapeutic applications.