Papers by William Blalock
Biomolecules
Since first being documented in ancient times, the relation of inflammation with injury and disea... more Since first being documented in ancient times, the relation of inflammation with injury and disease has evolved in complexity and causality. Early observations supported a cause (injury) and effect (inflammation) relationship, but the number of pathologies linked to chronic inflammation suggests that inflammation itself acts as a potent promoter of injury and disease. Additionally, results from studies over the last 25 years point to chronic inflammation and innate immune signaling as a critical link between stress (exogenous and endogenous) and adaptation. This brief review looks to highlight the role of the innate immune response in disease pathology, and recent findings indicating the innate immune response to chronic stresses as an influence in driving adaptation.
Biomolecules
Since first being documented in ancient times, the relation of inflammation with injury and disea... more Since first being documented in ancient times, the relation of inflammation with injury and disease has evolved in complexity and causality. Early observations supported a cause (injury) and effect (inflammation) relationship, but the number of pathologies linked to chronic inflammation suggests that inflammation itself acts as a potent promoter of injury and disease. Additionally, results from studies over the last 25 years point to chronic inflammation and innate immune signaling as a critical link between stress (exogenous and endogenous) and adaptation. This brief review looks to highlight the role of the innate immune response in disease pathology, and recent findings indicating the innate immune response to chronic stresses as an influence in driving adaptation.
Biomolecules
Since first being documented in ancient times, the relation of inflammation with injury and disea... more Since first being documented in ancient times, the relation of inflammation with injury and disease has evolved in complexity and causality. Early observations supported a cause (injury) and effect (inflammation) relationship, but the number of pathologies linked to chronic inflammation suggests that inflammation itself acts as a potent promoter of injury and disease. Additionally, results from studies over the last 25 years point to chronic inflammation and innate immune signaling as a critical link between stress (exogenous and endogenous) and adaptation. This brief review looks to highlight the role of the innate immune response in disease pathology, and recent findings indicating the innate immune response to chronic stresses as an influence in driving adaptation.
Identification of nuclear substrates of Akt/PKB by functional proteomics: prohibitin 2 is a target of Akt phosphorylation in human promyelocytic leukemia cells
The serine/threonine protein kinase Akt is a major signal transducer of the phosphoinositide 3-ki... more The serine/threonine protein kinase Akt is a major signal transducer of the phosphoinositide 3-kinase (PI 3-K) pathway in all cells and tissues and plays a pivotal role in the maintenance of cellular processes including cell growth, proliferation, survival, metabolism and development of many malignancies including acute myeloid leukemia. The frequent aberrant activation of the PI 3-K/Akt pathway in human cancer has made it an attractive therapeutic target. Therefore, the study of effector proteins downstream of Akt could clarify the role of Akt in the development of myeloid leukemia. Although both localization and activity of Akt in the nuclear compartment are well documented, most Akt substrates identified so far are located in the cytoplasm, while nuclear substrates have remained elusive. In this study, we applied a proteomic approach to identify novel Akt substrates by using an antibody that recognized a consensus motif phosphorylated by Akt (K/RXK/RXXS/T) when phosphorylated on ...
This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY
Ability of the activated PI3K/Akt oncoproteins to synergize with MEK1 and induce cell cycle progression and abrogate the cytokine-dependence of hematopoietic cells
Cell Cycle, Apr 1, 2004
Multiple signal transduction pathways, including the Raf/MEK/ERK and PI3K/Akt kinase cascades, pl... more Multiple signal transduction pathways, including the Raf/MEK/ERK and PI3K/Akt kinase cascades, play critical roles in transducing growth signals from activated cell surface receptors. Using conditionally and constitutively-active forms of MEK1 and either PI3K or Akt, we demonstrate synergy between these kinases in relieving cytokine-dependence of the FDC-P1 hematopoietic cell line. Cytokine-independent cells were obtained from DeltaMEK1:ER-infected cells at a frequency of 5 x 10(-5) indicating that low frequency of cells expressing beta-estradiol-regulated DeltaMEK1:ER became factor-independent, while activated PI3K or Akt by themselves did not relieve cytokine-dependence. In contrast, cytokine-independent cells were recovered approximately 25 to 250-fold more frequently from DeltaMEK1:ER infected cells also infected with either activated PI3K or Akt. MEK/PI3K and MEK/Akt-responsive cells could be maintained long-term as long as either beta-estradiol or the estrogen receptor antagon...
Elucidation of signal transduction pathways by transfection of cells with modified oncogenes
Msphere, 2003
This chapter will focus on introduction of various wild type (WT) and mutant genes into cells by ... more This chapter will focus on introduction of various wild type (WT) and mutant genes into cells by DNA transfection. Techniques for analysis of the inheritance, expression, and biological effects of the introduced genes will be described. Various strong and weak points about three different techniques of stable gene transfer, including calcium-phosphate DNA precipitation, transfection via liposomes, and transfection via electroporation, will be discussed.
Regulation of cell cycle progression and apoptosis by the Ras/Raf/MEK/ERK pathway (Review)
International Journal of Oncology, 2003
The Ras/Raf/MEK/ERK signal transduction pathway regulates cell cycle progression and apoptosis in... more The Ras/Raf/MEK/ERK signal transduction pathway regulates cell cycle progression and apoptosis in diverse types of cells. Mutations in this pathway are often observed in transformed cell lines and frequently linked with human cancers. The Ras/Raf/MEK/ERK pathway can induce events both associated with cell proliferation and cell cycle arrest. The particular course chosen may depend on the strength and the particular Raf gene activated by Ras. This pathway also is involved in maintaining cell survival by modulating the activity of apoptotic molecules including Bad and Bcl-2. This review will discuss the regulation of the Ras/Raf/MEK/ERK pathway and how it modulates cell cycle progression and cell survival.
![Research paper thumbnail of Proapoptotic activity and chemosensitizing effect of the novel Akt inhibitor (2S)-1-(1H-Indol-3-yl)-3-[5-(3methyl-2H-indazol-5-yl)pyridin-3-yl]oxypropan2-amine (A443654) in T acute lymphoblastic leukemia. Molecular Pharmacology](https://attachments.academia-assets.com/80676573/thumbnails/1.jpg)
Constitutively activated AKT kinase is a common feature of T-cell acute lymphoblastic leukemia (T... more Constitutively activated AKT kinase is a common feature of T-cell acute lymphoblastic leukemia (T-ALL). Here, we report that the novel AKT inhibitor (2S)-1-(1H-indol-3-yl)-3-[5-(3-methyl-2H-indazol-5-yl)pyridin-3-yl]oxypropan2-amine (A443654) leads to rapid cell death of T-ALL lines and patient samples. Treatment of CEM, Jurkat, and MOLT-4 cells with nanomolar doses of the inhibitor led to AKT phosphorylation accompanied by dephosphorylation and activation of the downstream target, glycogen synthase kinase-3β. Effects were time- and dose-dependent, resulting in apoptotic cell death. Treatment of Jurkat cells with A443654 resulted in activation of caspase-2,-3,-6,-8, and-9. Apoptotic cell death was mostly dependent on caspase-2 activation, as demonstrated by preincubation with a selective pharmacological inhibitor. It is remarkable that A443654 was highly effective against the drug-resistant cell line CEMVBL100, which expresses 170-kDa P-glycoprotein. Moreover, A443654 synergized wit...
Oncogene, 2000
1. Oncogene. 2000 Jan 27;19(4):526-36. A conditionally-active form of MEK1 results in autocrine t... more 1. Oncogene. 2000 Jan 27;19(4):526-36. A conditionally-active form of MEK1 results in autocrine tranformation of human and mouse hematopoietic cells. Blalock WL, Pearce M, Steelman LS, Franklin RA, McCarthy SA, Cherwinski H, McMahon M, McCubrey JA. ...
High levels of sIgA and exudated serum albumin in tears of contact lens related Dry Eye patients three months after discontinuation of lens use
Investigative Ophthalmology & Visual Science, 2013
Elucidation of signal transduction pathways by transfection of cells with modified oncogenes
Methods in molecular biology, 2003
This chapter will focus on introduction of various wild type (WT) and mutant genes into cells by ... more This chapter will focus on introduction of various wild type (WT) and mutant genes into cells by DNA transfection. Techniques for analysis of the inheritance, expression, and biological effects of the introduced genes will be described. Various strong and weak points about three different techniques of stable gene transfer, including calcium-phosphate DNA precipitation, transfection via liposomes, and transfection via electroporation, will be discussed.
Enhanced ability of daniplestim and mpo-1 to suppress apoptosis and stimulate cytokine-inducible gene expression
Blood, 2000
Abrogation of cytokine-dependency by MEK1 involves the PI3 kinase-AKT pathway and is enhanced by BCL2 overexpression or aberrant AKT activation
From the Dipartimento di Scienze Anatomiche Umane (F.F., W.L.B., F.C., L.C., and A.M.M.) and Dipa... more From the Dipartimento di Scienze Anatomiche Umane (F.F., W.L.B., F.C., L.C., and A.M.M.) and Dipartimento di Ematologia e Scienze Oncologiche “L. e A. Seràgnoli” (G.M.), Università di Bologna, Italy; Centro Immunoematologia e Trasfusionale, Policlinico S. Orsola-Malpighi, Bologna, Italy (P.L.T.); Dipartimento di Scienze Motorie e della Salute, Università di Cassino, Italy (A.C.); Dipartimento di Biotecnologie ed Ematologia, Università degli Studi “La Sapienza”, Roma, Italy (A.T.); Department of Microbiology and Immunology, Brody School of Medicine at East Carolina University, Greenville, NC, USA (J.A.M.) Molecular Pharmacology Fast Forward. Published on June 24, 2008 as doi:10.1124/mol.108.047639
Transformation of human hematopoietic cells by activated Raf oncoproteins Abilities of A-Raf and Raf-1 to induce MEK-dependent autocrine growth factor synthesis and inhibit differentiation
Rax-dependent PKR activation is required for host anti-viral defense after vesicular stomatitis virus infection
4450 The interferon-inducible dsRNA-dependent kinase PKR has been well studied as a component of ... more 4450 The interferon-inducible dsRNA-dependent kinase PKR has been well studied as a component of host anti-viral defense. In this context, PKR is activated by dsRNA byproducts produced during viral infection to phosphorylate the alpha subunit of eukaryotic initiation factor 2 (eIF2alpha) and stop de novo protein synthesis. In previous studies we discovered and characterized Rax, which to date is the only identified cellular PKR activator. Rax is also a dsRNA binding protein, yet the role of Rax in host antiviral defense is not clear. Now we report that the level of Rax expression in murine embryonic fibroblast (MEF) cells directly affects resistance to infection by vesicular stomatitis virus (VSV). Knock-down of Rax expression in MEF cells by 80% using siRNA prevents virus-induced PKR activation and eIF2alpha phosphorylation. Consequently, cells with reduced Rax display in creased VSV protein synthesis and viral titers >1000 fold higher than control cells 24 hours after infection...
RNA Processing and Ribosome Biogenesis in Bone Marrow Failure Disorders
RNA & DISEASE
Bone marrow failure disorders (BMFDs), which are characterized by an early pro-apoptotic phase wh... more Bone marrow failure disorders (BMFDs), which are characterized by an early pro-apoptotic phase which results in faulty hematopoiesis and anemia, more often than not progress to outright acute myelogenous leukemia (AML). Recent findings have indicated that most if not all of these disorders have a very significant RNA processing component to their pathology. This review aims to highlight some of normal processes of RNA metabolism that have been recently demonstrated to be altered in BMFDs.
The Raf/MEK/ERK kinase cascade is pivotal in transmitting signals from membrane receptors to tran... more The Raf/MEK/ERK kinase cascade is pivotal in transmitting signals from membrane receptors to transcription factors that control gene expression culminating in the regulation of cell cycle progression. This cascade can prevent cell death through ERK2 and p90 Rsk activation and phosphorylation of apoptotic and cell cycle regulatory proteins. The PI3K/Akt kinase cascade also controls apoptosis and can phosphorylate many apoptotic and cell cycle regulatory proteins. These pathways are interwoven as Akt can phosphorylate Raf and result in its inactivation, and Raf can be required for the antiapoptotic effects of Akt. In this study, the effects of activated Raf (Raf-1, A-Raf and B-Raf) and PI3K/Akt proteins on abrogation of cytokine dependence in FL5.12 hematopoietic cells were examined. Activated Raf, PI3K or Akt expression, by themselves, did not readily relieve cytokine dependence. The presence of activated Raf and PI3K/Akt increased the isolation of factor-independent cells from 400-to 2500-fold depending upon the particular combination examined. The individual effects of activated Raf and Akt on proliferation, apoptosis and autocrine growth factor synthesis were further examined with hormoneinducible constructs (DRaf-1:AR and DAkt:ER*(Myr +). Activation of either Raf or Akt hindered cell death; however, both proliferation and maximal synthesis of autocrine cytokines were dependent upon activation of both signaling pathways. The effects of small molecular weight inhibitors on DNA synthesis and cytokine gene expression were also examined. The PI3K inhibitor, LY294002, inhibited growth and cytokine gene expression. This effect could be synergistically increased by addition of the MEK inhibitor UO126. These cells will be useful in elucidating the interactions between Raf/MEK/ ERK and PI3K/Akt cascades in proliferation, apoptosis, and leukemogenesis, as well as evaluating the efficacy of signal transduction inhibitors that target these cascades.
The Raf/MEK/ERK kinase cascade plays a critical role in transducing growth signals from activated... more The Raf/MEK/ERK kinase cascade plays a critical role in transducing growth signals from activated cell surface receptors. Using DMEK1:ER, a conditionally active form of MEK1 which responds to either b-estradiol or the estrogen receptor antagonist 4 hydroxy-tamoxifen (4HT), we previously documented the ability of this dual specificity protein kinase to abrogate the cytokine-dependency of human (TF-1) and murine (FDC-P1 and FL5.12) hematopoietic cells lines. Here we demonstrate the ability of DMEK1:ER to activate the phosphatidylinositol 3kinase (PI3K)/Akt/p70 ribosomal S6 kinase (p70 S6K) pathway and the importance of this pathway in MEK1-mediated prevention of apoptosis. MEK1-responsive cells can be maintained long term in the presence of b-estradiol, 4HT or IL-3. Removal of hormone led to the rapid cessation of cell proliferation and the induction of apoptosis in a manner similar to cytokine deprivation of the parental cells. Stimulation of DMEK1:ER by 4HT resulted in ERK, PI3K, Akt and p70 S6K activation. Treatment with PI3K, Akt and p70 S6K inhibitors prevented MEK-responsive growth. Furthermore, the apoptotic effects of PI3K/Akt/p70 S6K inhibitors could be enhanced by cotreatment with MEK inhibitors. Use of a PI3K inhibitor and a constitutively active form of Akt, [DAkt(Myr +)], indicated that activation of PI3K was necessary for MEK1-responsive growth and survival as activation of Akt alone was unable to compensate for the loss of PI3K activity. Cells transduced by MEK or MEK+Akt displayed different sensitivities to signal transduction inhibitors, which targeted these pathways. These results indicate a requirement for the activation of the PI3K pathway during MEK-mediated transformation of certain hematopoietic cells. These experiments provide important clues as to why the identification of mutant signaling pathways may be the Achilles heel of leukemic cell growth. Leukemia treatment targeting multiple signal transduction pathways may be more efficacious than therapy aimed at inhibiting a single pathway.
Uploads
Papers by William Blalock