29 research outputs found
Health benefits attributed to 17α-estradiol, a lifespan-extending compound, are mediated through estrogen receptor α.
Metabolic dysfunction underlies several chronic diseases, many of which are exacerbated by obesity. Dietary interventions can reverse metabolic declines and slow aging, although compliance issues remain paramount. 17α-estradiol treatment improves metabolic parameters and slows aging in male mice. The mechanisms by which 17α-estradiol elicits these benefits remain unresolved. Herein, we show that 17α-estradiol elicits similar genomic binding and transcriptional activation through estrogen receptor α (ERα) to that of 17β-estradiol. In addition, we show that the ablation of ERα completely attenuates the beneficial metabolic effects of 17α-E2 in male mice. Our findings suggest that 17α-E2 may act through the liver and hypothalamus to improve metabolic parameters in male mice. Lastly, we also determined that 17α-E2 improves metabolic parameters in male rats, thereby proving that the beneficial effects of 17α-E2 are not limited to mice. Collectively, these studies suggest ERα may be a drug target for mitigating chronic diseases in male mammals
Hematopoietic reconstitution by multipotent adult progenitor cells: precursors to long-term hematopoietic stem cells
For decades, in vitro expansion of transplantable hematopoietic stem cells (HSCs) has been an elusive goal. Here, we demonstrate that multipotent adult progenitor cells (MAPCs), isolated from green fluorescent protein (GFP)-transgenic mice and expanded in vitro for >40–80 population doublings, are capable of multilineage hematopoietic engraftment of immunodeficient mice. Among MAPC-derived GFP+CD45.2+ cells in the bone marrow of engrafted mice, HSCs were present that could radioprotect and reconstitute multilineage hematopoiesis in secondary and tertiary recipients, as well as myeloid and lymphoid hematopoietic progenitor subsets and functional GFP+ MAPC-derived lymphocytes that were functional. Although hematopoietic contribution by MAPCs was comparable to control KTLS HSCs, approximately 103-fold more MAPCs were required for efficient engraftment. Because GFP+ host-derived CD45.1+ cells were not observed, fusion is not likely to account for the generation of HSCs by MAPCs
Reversal of hyperglycemia by insulin-secreting rat bone marrow- and blastocyst-derived hypoblast stem cell-like cells
β-cell replacement may efficiently cure type 1 diabetic (T1D) patients whose insulin-secreting β-cells have been selectively destroyed by autoantigen-reactive T cells. To generate insulin-secreting cells we used two cell sources: rat multipotent adult progenitor cells (rMAPC) and the highly similar rat extra-embryonic endoderm precursor (rXEN-P) cells isolated under rMAPC conditions from blastocysts (rHypoSC). rMAPC/rHypoSC were sequentially committed to definitive endoderm, pancreatic endoderm, and β-cell like cells. On day 21, 20% of rMAPC/rHypoSC progeny expressed Pdx1 and C-peptide. rMAPCr/HypoSC progeny secreted C-peptide under the stimulus of insulin agonist carbachol, and was inhibited by the L-type voltage-dependent calcium channel blocker nifedipine. When rMAPC or rHypoSC differentiated d21 progeny were grafted under the kidney capsule of streptozotocin-induced diabetic nude mice, hyperglycemia reversed after 4 weeks in 6/10 rMAPC- and 5/10 rHypoSC-transplanted mice. Hyperglycemia recurred within 24 hours of graft removal and the histological analysis of the retrieved grafts revealed presence of Pdx1-, Nkx6.1- and C-peptide-positive cells. The ability of both rMAPC and HypoSC to differentiate to functional β-cell like cells may serve to gain insight into signals that govern β-cell differentiation and aid in developing culture systems to commit other (pluripotent) stem cells to clinically useful β-cells for cell therapy of T1D
Multipotent adult progenitor cells sustain function of ischemic limbs in mice
Despite progress in cardiovascular research, a cure for peripheral vascular disease has not been found. We compared
the vascularization and tissue regeneration potential of murine and human undifferentiated multipotent
adult progenitor cells (mMAPC-U and hMAPC-U), murine MAPC-derived vascular progenitors (mMAPC-VP),
and unselected murine BM cells (mBMCs) in mice with moderate limb ischemia, reminiscent of intermittent
claudication in human patients. mMAPC-U durably restored blood flow and muscle function and stimulated
muscle regeneration, by direct and trophic contribution to vascular and skeletal muscle growth. This was in
contrast to mBMCs and mMAPC-VP, which did not affect muscle regeneration and provided only limited and
transient improvement. Moreover, mBMCs participated in a sustained inflammatory response in the lower
limb, associated with progressive deterioration in muscle function. Importantly, mMAPC-U and hMAPC-U also
remedied vascular and muscular deficiency in severe limb ischemia, representative of critical limb ischemia in
humans. Thus, unlike BMCs or vascular-committed progenitors, undifferentiated multipotent adult progenitor
cells offer the potential to durably repair ischemic damage in peripheral vascular disease patients
Multipotent adult progenitor cell transplantation increases vascularity and improves left ventricular function after myocardial infarction
Progressive contractile dysfunction of viable myocardium that surrounds a large infarct leads to
heart failure following acute myocardial infarction (AMI). Experimental evidence indicates that
cellular transplantation may improve the left ventricular (LV) contractile performance, even though
the underlying mechanisms remain undefined. Here, we compared the effect of transplantation of
murine multipotent adult progenitor cells (MAPCs), a population of adult bone marrow-derived
cells that differentiate into cells of mesodermal, endodermal and ectodermal origin, with murine
bone marrow cells (BMCs) or fibroblasts on post-infarct cardiac function by peri-infarct injection
after coronary artery ligation in mice. We demonstrate that, in contrast to the other cell populations,
transplantation of MAPCs significantly improved LV contractile function for at least 8 weeks posttransplantation
and, although BMCs reduced infarct size, the decrease in scar size was substantially
greater in MAPC-treated hearts. As neither MAPCs nor BMCs were present beyond 1 week, the
beneficial effect was not due to differentiation and direct contribution of MAPCs to the vascular
or cardiomyocyte compartment. Significantly more inflammatory cells were present in MAPC- than
BMC-treated hearts at 1 week, which was accompanied by increased vascularity 8 weeks posttransplantation.
We hypothesize that MAPCs indirectly contributed to these effects, by secreting
inflammatory [monocyte chemoattractant protein-1 (MCP)-1], and vascular growth factors [vascular
endothelial growth factor (VEGF), platelet-derived growth factor (PDGF)-BB, and transforming
growth factor (TGF)β1), and others, resulting in increased angiogenensis and cardioprotection
Reversal of hyperglycemia by insulin-secreting rat bone marrow- and blastocyst-derived hypoblast stem cell-like cells.
β-cell replacement may efficiently cure type 1 diabetic (T1D) patients whose insulin-secreting β-cells have been selectively destroyed by autoantigen-reactive T cells. To generate insulin-secreting cells we used two cell sources: rat multipotent adult progenitor cells (rMAPC) and the highly similar rat extra-embryonic endoderm precursor (rXEN-P) cells isolated under rMAPC conditions from blastocysts (rHypoSC). rMAPC/rHypoSC were sequentially committed to definitive endoderm, pancreatic endoderm, and β-cell like cells. On day 21, 20% of rMAPC/rHypoSC progeny expressed Pdx1 and C-peptide. rMAPCr/HypoSC progeny secreted C-peptide under the stimulus of insulin agonist carbachol, and was inhibited by the L-type voltage-dependent calcium channel blocker nifedipine. When rMAPC or rHypoSC differentiated d21 progeny were grafted under the kidney capsule of streptozotocin-induced diabetic nude mice, hyperglycemia reversed after 4 weeks in 6/10 rMAPC- and 5/10 rHypoSC-transplanted mice. Hyperglycemia recurred within 24 hours of graft removal and the histological analysis of the retrieved grafts revealed presence of Pdx1-, Nkx6.1- and C-peptide-positive cells. The ability of both rMAPC and HypoSC to differentiate to functional β-cell like cells may serve to gain insight into signals that govern β-cell differentiation and aid in developing culture systems to commit other (pluripotent) stem cells to clinically useful β-cells for cell therapy of T1D
Reversal of hyperglycemia by insulin-secreting rat bone marrow- and blastocyst-derived hypoblast stem cell-like cells
β-cell replacement may efficiently cure type 1 diabetic (T1D) patients whose insulin-secreting β-cells have been selectively destroyed by autoantigen-reactive T cells. To generate insulin-secreting cells we used two cell sources: rat multipotent adult progenitor cells (rMAPC) and the highly similar rat extra-embryonic endoderm precursor (rXEN-P) cells isolated under rMAPC conditions from blastocysts (rHypoSC). rMAPC/rHypoSC were sequentially committed to definitive endoderm, pancreatic endoderm, and β-cell like cells. On day 21, 20% of rMAPC/rHypoSC progeny expressed Pdx1 and C-peptide. rMAPCr/HypoSC progeny secreted C-peptide under the stimulus of insulin agonist carbachol, and was inhibited by the L-type voltage-dependent calcium channel blocker nifedipine. When rMAPC or rHypoSC differentiated d21 progeny were grafted under the kidney capsule of streptozotocin-induced diabetic nude mice, hyperglycemia reversed after 4 weeks in 6/10 rMAPC- and 5/10 rHypoSC-transplanted mice. Hyperglycemia recurred within 24 hours of graft removal and the histological analysis of the retrieved grafts revealed presence of Pdx1-, Nkx6.1- and C-peptide-positive cells. The ability of both rMAPC and HypoSC to differentiate to functional β-cell like cells may serve to gain insight into signals that govern β-cell differentiation and aid in developing culture systems to commit other (pluripotent) stem cells to clinically useful β-cells for cell therapy of T1D
High-Throughput Screening for Growth Inhibitors Using a Yeast Model of Familial Paraganglioma
<div><p>Classical tumor suppressor genes block neoplasia by regulating cell growth and death. A remarkable puzzle is therefore presented by familial paraganglioma (PGL), a neuroendocrine cancer where the tumor suppressor genes encode subunits of succinate dehydrogenase (SDH), an enzyme of the tricarboxylic acid (TCA) cycle of central metabolism. Loss of SDH initiates PGL through mechanisms that remain unclear. Could this metabolic defect provide a novel opportunity for chemotherapy of PGL? We report the results of high throughput screening to identify compounds differentially toxic to SDH mutant cells using a powerful <i>S. cerevisiae</i> (yeast) model of PGL. Screening more than 200,000 compounds identifies 12 compounds that are differentially toxic to SDH-mutant yeast. Interestingly, two of the agents, dequalinium and tetraethylthiuram disulfide (disulfiram), are anti-malarials with the latter reported to be a glycolysis inhibitor. We show that four of the additional hits are potent inhibitors of yeast alcohol dehydrogenase. Because alcohol dehydrogenase regenerates NAD<sup>+</sup> in glycolytic cells that lack TCA cycle function, this result raises the possibility that lactate dehydrogenase, which plays the equivalent role in human cells, might be a target of interest for PGL therapy. We confirm that human cells deficient in SDH are differentially sensitive to a lactate dehydrogenase inhibitor.</p> </div
Thymidine Analogs Are Transferred from Prelabeled Donor to Host Cells in the Central Nervous System After Transplantation: A Word of Caution
Thymidine analogs, including bromodeoxyuridine, chlorodeoxyuridine,
iododeoxyuridine, and tritiated thymidine, label
dividing cells by incorporating into DNA during S phase of cell
division and are widely employed to identify cells transplanted
into the central nervous system. However, the potential for
transfer of thymidine analogs from grafted cells to dividing
host cells has not been thoroughly tested. We here demonstrate
that graft-derived thymidine analogs can become incorporated
into host neural precursors and glia. Large numbers of labeled
neurons and glia were found 3–12 weeks after transplantation
of thymidine analog-labeled live stem cells, suggesting differentiation
of grafted cells. Remarkably, however, similar results
were obtained after transplantation of dead cells or labeled
fibroblasts. Our findings reveal for the first time that thymidine
analog labeling may not be a reliable means of identifying
transplanted cells, particularly in highly proliferative environments
such as the developing, neurogenic, or injured brain