X-Message-Number: 27429 Date: Wed, 14 Dec 2005 21:33:08 -0800 (PST) From: Doug Skrecky <> Subject: GSK-3 inhibitors offer a variety of benefits [An argument could be made in favor of taking lithium, as a GSK-3 inhibitor for life-extension purposes.] Nat Med 2005 Dec 11; [Epub ahead of print Glycogen synthase kinase-3 is an in vivo regulator of h ematopoietic stem cell repopulation. The in vivo regulation of hematopoietic stem cell (HSC) function is poorly understood. Here, we show that hematopoietic repopulation can be augmented by administration of a glycogen synthase kinase-3 (GSK-3) inhibitor to recipient mice transplanted with mouse or human HSCs. GSK-3 inhibitor treatment improved neutrophil and megakaryocyte recovery, recipient survival and resulted in enhanced sustained long-term repopulation. The output of primitive Lin(-)c-Kit(+)Sca-1(+) cells and progenitors from HSCs increased upon GSK-3 inhibitor treatment without altering secondary repopulating ability, suggesting that the HSC pool is maintained while overall hematopoietic reconstitution is increased. GSK-3 inhibitors were found to modulate gene targets of Wnt, Hedgehog and Notch pathways in cells comprising the primitive hematopoietic compartment without affecting mature cells. Our study establishes GSK-3 as a specific in vivo modulator of HSC activity, and suggests that administration of GSK-3 inhibitors may provide a clinical means to directly enhance the repopulating capacity of transplanted HSCs. Circulation. 2005 Aug 30;112(9):1316-22. Glycogen synthase kinase-3 mediates endothelial cell activation by tumor necrosis factor-alpha. BACKGROUND: Endothelial cell transformation to a thrombogenic and inflammatory phenotype plays an important role in the pathogenesis of atherothrombosis, but the responsible signaling pathways remain to be elucidated. This study was designed to investigate the regulatory role of glycogen synthase kinase-3 (GSK-3) in tissue factor (TF) and vascular cell adhesion molecule (VCAM)-1 expression in tumor necrosis factor (TNF)-alpha-stimulated endothelial cells. METHODS AND RESULTS: In human endothelial cells, TNF-alpha as well as thrombin induced rapid and transient dephosphorylation and hence, activation of GSK-3. A GSK-3 inhibitor, LiCl, suppressed TNF-alpha- and thrombin-induced TF and VCAM-1 expression, whereas NaCl had no effect. A specific GSK-3 inhibitor, TDZD-8, mimicked the inhibitory effects of lithium. GSK-3 inhibition also significantly suppressed the TNF-alpha-induced increase in TF activity and VCAM-1 cell-surface expression. The luciferase reporter system demonstrated that regulation of TF and VCAM-1 expression by GSK-3 was mediated at the transcriptional level. The TNF-alpha-induced increase in nuclear factor (NF)-kappaB DNA-binding activity was significantly suppressed by TDZD-8. TDZD-8 completely prevented the TNF-alpha-induced inhibitor of NF-kappaB (IkappaB)-alpha degradation but had no effect on IkappaB-kinase-beta phosphorylation. CONCLUSIONS: GSK-3 regulates TNF-alpha-induced IkappaB-alpha degradation and NF-kappaB activation independent of IkappaB-kinase-beta and subsequent induction of TF and VCAM-1 expression in human endothelial cells. This study provides the experimental basis for a novel strategy of using GSK-3 inhibition to treat atherothrombotic vascular disease. Cancer Res. 2005 Apr 1;65(7):2537-41. Valproic acid stimulates proliferation and self-renewal of hematopoietic stem cells. Histone deacetylase inhibitors have attracted considerable attention because of their ability to overcome the differentiation block in leukemic blasts, an effect achieved either alone or in combination with differentiating agents, such as all-trans retinoic acid. We have previously reported favorable effects of the potent histone deacetylase inhibitor valproic acid in combination with all-trans retinoic acid in patients with advanced acute myeloid leukemia leading to blast cell reduction and improvement of hemoglobin. These effects were accompanied by hypergranulocytosis most likely due to an enhancement of nonleukemic myelopoiesis and the suppression of malignant hematopoiesis rather than enforced differentiation of the leukemic cells. These data prompted us to investigate the effect of valproic acid on normal hematopoietic stem cells (HSC). Here we show that valproic acid increases both proliferation and self-renewal of HSC. It accelerates cell cycle progression of HSC accompanied by a down-regulation of p21(cip-1/waf-1). Furthermore, valproic acid inhibits GSK3beta by phosphorylation on Ser9 accompanied by an activation of the Wnt signaling pathway as well as by an up-regulation of HoxB4, a target gene of Wnt signaling. Both are known to directly stimulate the proliferation of HSC and to expand the HSC pool. In summary, we here show that valproic acid, known to induce differentiation or apoptosis in leukemic blasts, stimulates the proliferation of normal HSC, an effect with a potential effect on its future role in the treatment of acute myeloid leukemia. Neuropharmacology. 2005 Mar;48(4):576-83. Epub 2005 Jan 25. Opposite effects of lithium and valproic acid on trophic factor deprivation-induced glycogen synthase kinase-3 activation, c-Jun expression and neuronal cell death. Recent studies demonstrate that lithium and valproic acid (VPA), two commonly used mood-stabilizing drugs, have neuroprotective effects against a variety of insults. Inhibition of the proapoptotic enzyme, glycogen synthase kinase-3 (GSK-3), has been suggested to be the mechanism of action of neuroprotection for both drugs. In this study, we tested if lithium and VPA could protect cultured cerebellar granule neurons (CGNs) from GSK-3-mediated apoptosis induced by trophic factor withdrawal (serum/potassium deprivation). Both lithium and indirubin, a specific GSK-3 inhibitor, protected CGNs in a dose-dependent manner. In contrast, VPA did not provide any neuroprotection and even potentiated cell death. Immunoblot analysis revealed that lithium inhibited the trophic factor deprivation-induced activation of GSK-3 as well as the in vivo phosphorylation of the microtubule-associated protein Tau on Ser199, a specific target site for GSK-3. Under these same experimental conditions, however, VPA neither inhibited GSK-3 activation nor hindered GSK-3 mediated Tau phosphorylation. Furthermore, in accordance with their effects on neuronal survival, lithium prevented the induction of c-Jun expression in trophic factor-deprived CGNs, whereas VPA potentiated it. Collectively, these results show that VPA is not a universal inhibitor of neuronal GSK-3, and that instead of being neuroprotective, VPA can even exacerbate neuronal death under some conditions. Farmakol Toksikol. 1986 May-Jun;49(3):69-70. [Effect of lithium oxybutyrate on the viability of preserved donor kidney] The experiments on 10 dogs showed that replacement of sodium oxybutyrate for lithium oxybutyrate in the solution for preservation increases the period of preserving the structural and functional integrity of the donor kidney, contributes to its survival and delays the development of the transplant rejection. 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