DOCSLIB.ORG

Anaplastic Lymphoma Kinase (ALK): Structure, Oncogenic Activation, and Pharmacological Inhibition

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Anaplastic Lymphoma Kinase (ALK): Structure, Oncogenic Activation, and Pharmacological Inhibition Pharmacological Research 68 (2013) 68–94 Contents lists available at SciVerse ScienceDirect Pharmacological Research jo urnal homepage: www.elsevier.com/locate/yphrs Invited review Anaplastic lymphoma kinase (ALK): Structure, oncogenic activation, and pharmacological inhibition ∗ Robert Roskoski Jr. Blue Ridge Institute for Medical Research, 3754 Brevard Road, Suite 116, Box 19, Horse Shoe, NC 28742, USA a r t i c l e i n f o a b s t r a c t Article history: Anaplastic lymphoma kinase was first described in 1994 as the NPM-ALK fusion protein that is expressed Received 14 November 2012 in the majority of anaplastic large-cell lymphomas. ALK is a receptor protein-tyrosine kinase that was Accepted 18 November 2012 more fully characterized in 1997. Physiological ALK participates in embryonic nervous system develop- ment, but its expression decreases after birth. ALK is a member of the insulin receptor superfamily and Keywords: is most closely related to leukocyte tyrosine kinase (Ltk), which is a receptor protein-tyrosine kinase. Crizotinib Twenty different ALK-fusion proteins have been described that result from various chromosomal rear- Drug discovery rangements, and they have been implicated in the pathogenesis of several diseases including anaplastic Non-small cell lung cancer large-cell lymphoma, diffuse large B-cell lymphoma, and inflammatory myofibroblastic tumors. The Protein kinase inhibitor EML4-ALK fusion protein and four other ALK-fusion proteins play a fundamental role in the development Targeted cancer therapy Acquired drug resistance in about 5% of non-small cell lung cancers. The formation of dimers by the amino-terminal portion of the ALK fusion proteins results in the activation of the ALK protein kinase domain that plays a key role in the tumorigenic process. Downstream signaling from ALK fusion proteins involves the Ras/Raf/MEK/ERK1/2 cell proliferation module and the JAK/STAT cell survival pathway. Furthermore, nearly two dozen ALK activating mutations participate in the pathogenesis of childhood neuroblastomas along with ALK over- expression. The occurrence of oncogenic ALK, particularly in non-small cell lung cancer, has generated considerable interest and effort in developing ALK inhibitors. Currently, crizotinib has been approved by the US Food and Drug Administration for the treatment of ALK-positive non-small cell lung cancer along with an approved fluorescence in situ hybridization kit used for the diagnosis of the disease. The emergence of crizotinib drug resistance with a median occurrence at approximately 10 months after the initiation of therapy has stimulated the development of second-generation drugs for the treatment of non-small cell lung cancer and other disorders. About 28% of the cases of crizotinib resistance are related to nearly a dozen different mutations of ALK in the EML4-ALK fusion protein; the other cases of resis- tance are related to the upregulation of alternative signaling pathways or to undefined mechanisms. It is remarkable that the EML4-ALK fusion protein was discovered in 2007 and crizotinib was approved for the treatment of ALK-positive non-small cell lung cancer in 2011, which is a remarkably short timeframe in the overall scheme of drug discovery. © 2012 Elsevier Ltd. All rights reserved. Contents 1. Introduction . 69 2. An overview of anaplastic lymphoma kinase (ALK) . 70 3. Structure of the ALK protein kinase domain. 71 3.1. Catalytic residues in the amino- and carboxyterminal lobes of the ALK protein kinase domain . 71 3.2. ALK hydrophobic spines . 73 Abbreviations: AL, activation loop; ALCL, anaplastic large-cell lymphoma; C-spine, catalytic spine; CRC, colorectal cancer; D, distribution coefficient; DLBCL, diffuse large B-cell lymphoma; EGFR, epidermal growth factor receptor; HGF/SF, hepatocyte growth factor/scatter factor; IMT, inflammatory myofibroblastic tumor; IRS1, insulin receptor substrate 1; JAK, Janus activated kinase; LE, ligand efficiency; LipE, lipophilic efficiency; MW, molecular weight; NSCLC, non-small cell lung cancer; P, partition coefficient; PDGFR, platelet-derived growth factor receptor; PI3K, phosphatidylinositol 3-kinase; PKA, protein kinase A; PKC, protein kinase C; PLC, phospholipase C; pY, phosphotyrosine; R-spine, regulatory spine; VEGFR, vascular endothelial growth factor receptor. ∗ Tel.: +1 828 891 5637; fax: +1 828 890 8130. E-mail address: [email protected] 1043-6618/$ – see front matter © 2012 Elsevier Ltd. All rights reserved. http://dx.doi.org/10.1016/j.phrs.2012.11.007 R. Roskoski Jr. / Pharmacological Research 68 (2013) 68–94 69 3.3. Structure of quiescent ALK protein kinase . 73 3.4. Steady-state enzyme kinetic parameters of quiescent and active ALK . 74 4. ALK phosphorylation, activation, and downstream signaling . 74 4.1. ALK phosphorylation and activation . 74 4.2. ALK signaling . 74 5. ALK in disease: fusion proteins, mutants, and overexpression. 75 5.1. Anaplastic large-cell lymphoma . 75 5.2. Non-small cell lung cancer . 76 5.3. Diffuse large B-cell lymphoma . 76 5.4. Inflammatory myofibroblastic tumors . 77 5.5. Neuroblastoma . 78 5.6. Anaplastic thyroid cancer . 78 5.7. Rhabdomyosarcoma . 78 6. Oncogenic activation of ALK . 79 6.1. ALK activation following chromosomal translocations or inversions with the formation of ALK fusion proteins . 79 6.2. ALK activation by missense mutations in neuroblastomas and in anaplastic thyroid carcinomas . 79 7. ALK inhibitors ... 81 7.1. Crizotinib: a combined ALK and c-Met inhibitor . 81 7.1.1. ALK and c-Met as drug targets . 81 7.1.2. Development of crizotinib . 81 7.1.3. Inhibition of tumor growth and drug-induced toxicity . 82 7.1.4. Acquired crizotinib resistance . 83 7.2. CH5424842 . ..
Load more

Recommended publications
  • AMPK Exerts Dual Regulatory Effects on the PI3K Pathway Rong Tao1, Jun Gong2, Xixi Luo3, Mengwei Zang4, Wen Guo4, Rong Wen5, Zhijun Luo2,4*
    Tao et al. Journal of Molecular Signaling 2010, 5:1 http://www.jmolecularsignaling.com/content/5/1/1 RESEARCH ARTICLE Open Access AMPK exerts dual regulatory effects on the PI3K pathway Rong Tao1, Jun Gong2, Xixi Luo3, Mengwei Zang4, Wen Guo4, Rong Wen5, Zhijun Luo2,4* Abstract Background: AMP-activated protein kinase (AMPK) is a fuel-sensing enzyme that is activated when cells experience energy deficiency and conversely suppressed in surfeit of energy supply. AMPK activation improves insulin sensitivity via multiple mechanisms, among which AMPK suppresses mTOR/S6K-mediated negative feedback regulation of insulin signaling. Results: In the present study we further investigated the mechanism of AMPK-regulated insulin signaling. Our results showed that 5-aminoimidazole-4-carboxamide-1 ribonucleoside (AICAR) greatly enhanced the ability of insulin to stimulate the insulin receptor substrate-1 (IRS1)-associated PI3K activity in differentiated 3T3-F442a adipocytes, leading to increased Akt phosphorylation at S473, whereas insulin-stimulated activation of mTOR was diminished. In 3T3-F442a preadipocytes, these effects were attenuated by expression of a dominant negative mutant of AMPK a1 subunit. The enhancing effect of ACIAR on Akt phosphorylation was also observed when the cells were treated with EGF, suggesting that it is regulated at a step beyond IR/IRS1. Indeed, when the cells were chronically treated with AICAR in the absence of insulin, Akt phosphorylation was progressively increased. This event was associated with an increase in levels of phosphatidylinositol -3,4,5-trisphosphate (PIP3) and blocked by Wortmannin. We then expressed the dominant negative mutant of PTEN (C124S) and found that the inhibition of endogenous PTEN per se did not affect phosphorylation of Akt at basal levels or upon treatment with AICAR or insulin.
    [Show full text]
  • 2025.Full-Text.Pdf
    Human Cancer Biology Inhibition of Phosphotyrosine Phosphatase 1B Causes Resistance in BCR-ABL-PositiveLeukemiaCellstotheABLKinaseInhibitorSTI571 Noriko Koyama,1Steffen Koschmieder,1SandhyaTyagi,1Ignacio Portero-Robles,1Jo« rg Chromic,1 Silke Myloch,1Heike Nu« rnberger,1Ta nja Ro s s m a ni t h , 1Wolf-Karsten Hofmann,2 Dieter Hoelzer,1and Oliver Gerhard Ottmann1 Abstract Protein tyrosine phosphatase 1B(PTP1B) is a negative regulator of BCR-ABL-mediated transfor- mation in vitro and in vivo. Toinvestigate whether PTP1B modulates the biological effects of the abl kinase inhibitor STI571in BCR-ABL-positive cells, we transfected Philadelphia chromosome ^ positive (Ph+) chronic myeloid leukemia cell-derived K562 cells with either wild-type PTP1B (K562/PTP1B), a substrate-trapping dominant-negative mutant PTP1B(K562/D181A), or empty vector (K562/mock). Cells were cultured with or without STI571and analyzed for its effects on proliferation, differentiation, and apoptosis. In both K562/mock and K562/PTP1B cells, 0.25 to 1 Amol/L STI571 induced dose-dependent growth arrest and apoptosis, as measured by a decrease of cell proliferation and an increase of Annexin V-positive cells and/or of cells in the sub-G1 apoptotic phase. Western blot analysis showed increased protein levels of activated caspase-3 and caspase-8 and induction of poly(ADP-ribose) polymerase cleavage. Low con- centrations of STI571promoted erythroid differentiation of these cells. Conversely, K562/D181A cells displayed significantly lower PTP1B-specific tyrosine phosphatase activity and were signifi- cantly less sensitive to STI571-induced growth arrest, apoptosis, and erythroid differentiation. Pharmacologic inhibition of PTP1B activity in wild-type K562 cells, using bis(N,N-dimethylhy- droxamido)hydroxooxovanadate, attenuated STI571-induced apoptosis.
    [Show full text]
  • NATIONAL INSTITUTE for HEALTH and CLINICAL EXCELLENCE Final Appraisal Determination Erlotinib for the Treatment of Non-Small-Cell Lung Cancer
    CONFIDENTIAL NATIONAL INSTITUTE FOR HEALTH AND CLINICAL EXCELLENCE Final appraisal determination Erlotinib for the treatment of non-small-cell lung cancer This guidance was developed using the single technology appraisal (STA) process. 1 Guidance 1.1 Erlotinib is recommended, within its licensed indication, as an alternative to docetaxel as a second-line treatment option for patients with non-small-cell lung cancer (NSCLC) only on the basis that it is provided by the manufacturer at an overall treatment cost (including administration, adverse events and monitoring costs) equal to that of docetaxel. 1.2 The decision to use erlotinib or docetaxel (as outlined in section 1.1) should be made after a discussion between the responsible clinician and the individual about the potential benefits and adverse effects of each treatment. 1.3 Erlotinib is not recommended for the second-line treatment of locally advanced or metastatic NSCLC in patients for whom docetaxel is unsuitable (that is, where there is intolerance of or contraindications to docetaxel) or for third-line treatment after docetaxel therapy. 1.4 People currently receiving treatment with erlotinib, but for whom treatment would not be recommended according to section 1.3, should have the option to continue treatment until they and their clinicians consider it appropriate to stop. National Institute for Health and Clinical Excellence Page 1 of 29 Final appraisal determination – Erlotinib for the treatment of non-small-cell lung cancer Issue date: September 2008 CONFIDENTIAL 2 The technology 2.1 Erlotinib (Tarceva, Roche Products) is an orally active inhibitor of the epidermal growth factor receptor (EGFR) tyrosine kinase.
    [Show full text]
  • Insulin Receptor Substrates-5 and -6 Are Poor Substrates for the Insulin Receptor
    MOLECULAR MEDICINE REPORTS 3: 189-193, 2010 189 Insulin receptor substrates-5 and -6 are poor substrates for the insulin receptor SoETkIn VERSTEyHE1, CHRISToPHE BlanqUart2,3, CoRnElIa HamPE2,3, SHaUkaT maHmooD1, nEVEna CHRISTEFF2,3, PIERRE DE MEYTS1, STEVEn G. GRay1,4 and TARIK ISSAD2,3 1Receptor Systems Biology laboratory, Hagedorn Research Institute, novo nordisk a/S, 2820 Gentofte, Denmark; 2Institut Cochin, Université Paris Descartes, CnRS (UmR 8104), 75014 Paris; 3Inserm, U567, 75654 Paris, France; 4Translational Cancer Research Group, Trinity Centre for Health Sciences, Institute of molecular medicine, St James's Hospital, Dublin 8, Ireland Received September 30, 2009; accepted november 13, 2009 DoI: 10.3892/mmr_00000239 Abstract. Insulin receptor substrates (IRS)-5 and -6 are two leads to the activation of receptor tyrosine kinase and receptor recently identified members of the IRS family. We investi- phosphorylation, which enables the binding of docking proteins gated their roles as insulin receptor substrates and compared such as insulin receptor substrates (IRS)-1, -2, -3 and -4 and them with Src-homology-2-containing (Shc) protein, a Src-homology-2-containing protein (Shc). This in turn leads to well-established substrate. Bioluminescence resonance their phosphorylation and, thereby, intracellular signalling (1). energy transfer (BRET) experiments showed no interaction Until recently, the IRS family included IRS-1, -2, -3 and between the receptor and IRS-5, while interaction with IRS-6 -4 (2) and three downstream of kinases, Dok-1, -2 and -3. was not enhanced by insulin. By contrast, Shc showed an These seven proteins have similar amino-terminal pleckstrin insulin-induced BRET response, as did a truncated form of homology (PH) and similar phosphotyrosine binding (PTB) IRS-1 (1-262).
    [Show full text]
  • DNA Damage and the Activation of the P53 Pathway Mediate Alterations in Metabolic and Secretory Functions of Adipocytes
    3062 Diabetes Volume 65, October 2016 Bastien Vergoni,1,2 Pierre-Jean Cornejo,1,2 Jérôme Gilleron,1,2 Mansour Djedaini,1,2 Franck Ceppo,1,2 Arnaud Jacquel,2,3 Gwennaelle Bouget,1,2 Clémence Ginet,1,2 Teresa Gonzalez,1,2,4 Julie Maillet,5,6,7 Véronique Dhennin,5,6,7 Marie Verbanck,5,6,7 Patrick Auberger,2,3 Philippe Froguel,5,6,7,8 Jean-François Tanti,1,2 and Mireille Cormont1,2 DNA Damage and the Activation of the p53 Pathway Mediate Alterations in Metabolic and Secretory Functions of Adipocytes Diabetes 2016;65:3062–3074 | DOI: 10.2337/db16-0014 Activation of the p53 pathway in adipose tissue contributes Insulin resistance is associated with obesity and is a major to insulin resistance associated with obesity. However, the risk factor for type 2 diabetes. Adipose tissue (AT) plays mechanisms of p53 activation and the effect on adipocyte an important role in glucose and lipid homeostasis (1,2), functions are still elusive. Here we found a higher level of and AT dysfunction associated with obesity has emerged DNA oxidation and a reduction in telomere length in adipose as a critical event for the development of insulin resis- tissueofmicefedahigh-fatdietandanincreaseinDNA tance. Low-grade inflammation and oxidative and hypoxic damage and activation of the p53 pathway in adipocytes. stresses both develop in obese AT and are involved in the Interestingly, hallmarks of chronic DNA damage are visible alteration of adipocyte functions (3–6). Therefore, activa- at the onset of obesity. Furthermore, injection of lean mice tion of stress-signaling pathways in adipocytes is critically with doxorubicin, a DNA damage-inducing drug, increased involved in AT dysfunction and insulin resistance (7–10).
    [Show full text]
  • Insulin Activates the Insulin Receptor to Downregulate the PTEN Tumour Suppressor
    Oncogene (2014) 33, 3878–3885 OPEN & 2014 Macmillan Publishers Limited All rights reserved 0950-9232/14 www.nature.com/onc ORIGINAL ARTICLE Insulin activates the insulin receptor to downregulate the PTEN tumour suppressor J Liu1, S Visser-Grieve2,4, J Boudreau1, B Yeung2,SLo1, G Chamberlain1,FYu1, T Sun3,5, T Papanicolaou1, A Lam1, X Yang2 and I Chin-Sang1 Insulin and insulin-like growth factor-1 signaling have fundamental roles in energy metabolism, growth and development. Recent research suggests hyperactive insulin receptor (IR) and hyperinsulinemia are cancer risk factors. However, the mechanisms that account for the link between the hyperactive insulin signaling and cancer risk are not well understood. Here we show that an insulin-like signaling inhibits the DAF-18/(phosphatase and tensin homolog) PTEN tumour suppressor in Caenorhabditis elegans and that this regulation is conserved in human breast cancer cells. We show that inhibiting the IR increases PTEN protein levels, while increasing insulin signaling decreases PTEN protein levels. Our results show that the kinase region of IRb subunit physically binds to PTEN and phosphorylates on Y27 and Y174. Our genetic results also show that DAF-2/IR negatively regulates DAF-18/PTEN during C. elegans axon guidance. As PTEN is an important tumour suppressor, our results therefore suggest a possible mechanism for increased cancer risk observed in hyperinsulinemia and hyperactive IR individuals. Oncogene (2014) 33, 3878–3885; doi:10.1038/onc.2013.347; published online 2 September 2013 Keywords: PTEN; C. elegans; tumour suppressor; cancer; insulin signaling; genetics INTRODUCTION tumour suppression and further substantiate the importance of Insulin and insulin-like growth factor (IGF)-1 signaling are the PTEN regulation in cancer progression.
    [Show full text]
  • Treatment Paradigms in Advanced Non–Small-Cell Lung Cancer Caroline E
    Treatment Paradigms in Advanced Non–Small-Cell Lung Cancer Caroline E. McCoach, MD, PhD, and Karen Kelly, MD Dr McCoach is a clinical research fellow Abstract: Lung cancer is the most common cause of cancer-relat- and Dr Kelly is a professor of medicine, ed death worldwide, owing to its metastatic spread at the time of an associate director for clinical research, diagnosis. As a result, chemotherapy is the standard of care for the and the Jennifer Rene Harmon Tegley majority of patients. In recent years, the role of chemotherapy has and Elizabeth Erica Harmon Endowed Chair in Cancer Clinical Research in the expanded to include maintenance therapy and approved second- department of internal medicine at the and third-line treatments. Nonetheless, traditional chemotherapy University of California, Davis Medical has modestly improved outcomes in patients with advanced Center in Sacramento, California. non–small-cell lung cancer (NSCLC). Research efforts have been redirected toward the integration of molecularly-targeted agents Address correspondence to: into a treatment algorithm with unprecedented survival rates Karen Kelly, MD UC Davis Comprehensive Cancer Center in selected patients. This article will provide an update on the 4501 X Street, Suite 3016 multiple systemic regimens available to treat NSCLC, and discuss Sacramento, CA 95817 emerging molecular-based therapies. Phone: 916-734-3735 Fax: 916-734-7946 E-mail: [email protected] Introduction Lung cancer is the leading cause of cancer-related death worldwide, contributing to an estimated 1.4 million deaths every year.1 This high mortality rate results from the inability to detect lung cancer in its early stage.
    [Show full text]
  • Selective Insulin Signaling Through a and B Insulin Receptors Regulates Transcription of Insulin and Glucokinase Genes in Pancreatic ␤ Cells
    Molecular Cell, Vol. 7, 559±570, March, 2001, Copyright 2001 by Cell Press Selective Insulin Signaling through A and B Insulin Receptors Regulates Transcription of Insulin and Glucokinase Genes in Pancreatic ␤ Cells Barbara Leibiger,*§ Ingo B. Leibiger,*§k ceptors as the primary target, include signaling via mito- Tilo Moede,* Sabine Kemper,* gen-activated protein (MAP) kinases and phosphoinosi- Rohit N. Kulkarni,² C. Ronald Kahn,² tol-3 kinase (PI3K). The insulin receptor (IR), the first Lina Moitoso de Vargas,³ and Per-Olof Berggren* step in these cascades, exists in two isoforms as a result *The Rolf Luft Center for Diabetes Research of alternative mRNA splicing of the 11th exon of the insulin Department of Molecular Medicine proreceptor transcript (Seino et al., 1989). The A type Karolinska Institutet (IR-A), or Ex11Ϫ (Ullrich et al., 1985), lacks whereas the S-171 76 Stockholm B type (IR-B), or Ex11ϩ (Ebina et al., 1985), contains Sweden the respective sequence coding for 12 amino acids in ² Research Division the C terminus of the ␣ chain of the receptor. To date, Joslin Diabetes Center and no insulin-induced effect has been reported that dis- Department of Medicine criminates signaling via A- and B-type receptors. In fact, Harvard Medical School the functional significance of these IR isoforms remains Boston, Massachusetts 02215 unclear. ³ Department of Medicine Recent studies have shown that the insulin-producing New England Medical Center and pancreatic ␤ cell is a target for insulin action, with insulin Tufts University School of Medicine effects on transcription, translation, Ca2ϩ flux, and exo- Boston, Massachusetts 02111 cytosis (Leibiger et al., 1998a, 2000; Xu and Rothenberg, 1998; Xu et al., 1998; Aspinwall et al., 1999; Kulkarni et al., 1999a).
    [Show full text]
  • Inhibition of Insulin Receptor Gene Expression and Insulin Signaling by Fatty Acid: Interplay of PKC Isoforms Therein
    Original Paper Cellular Physiology Cell Physiol Biochem 2005;16:217-228 Accepted: July 27, 2005 and Biochemistry Inhibition of Insulin Receptor Gene Expression and Insulin Signaling by Fatty Acid: Interplay of PKC Isoforms Therein Debleena Dey, Mohua Mukherjee, Dipanjan Basu1, Malabika Datta, Sib Sankar Roy, Arun Bandyopadhyay and Samir Bhattacharya1 Molecular Endocrinology Laboratory, Indian Institute of Chemical Biology, 4, Raja S. C. Mullick Road, Kolkata, 1Cellular and Molecular Endocrinology Laboratory, Department of Zoology, School of Life Science, Visva-Bharati University, Santiniketan Key Words dependent, palmitate effected its constitutive Insulin resistance • Type 2 diabetes • Insulin receptor phosphorylation independent of PDK1. Time kinetics • Insulin signaling • PKC isoforms • Free fatty acids • study showed translocation of palmitate induced HMG phosphorylated PKCε from cell membrane to nuclear region and its possible association with the inhibition Abstract of IR gene transcription. Our study suggests one of Fatty acids are known to play a key role in promoting the pathways through which fatty acid can induce the loss of insulin sensitivity causing insulin resistance insulin resistance in skeletal muscle cell. and type 2 diabetes. However, underlying mechanism involved here is still unclear. Incubation of rat skeletal muscle cells with palmitate followed by I125- insulin Copyright © 2005 S. Karger AG, Basel binding to the plasma membrane receptor preparation demonstrated a two-fold decrease in receptor occupation. In searching the cause for this reduction, Introduction we found that palmitate inhibition of insulin receptor (IR) gene expression effecting reduced amount of IR Insulin resistance and type 2 diabetes mellitus is an protein in skeletal muscle cells. This was followed by insidious disease that accounts for more than 95% of the inhibition of insulin-stimulated IRβ tyrosine diabetic cases.
    [Show full text]
  • Real-Time Quantification of BCR-ABL Mrna Transcripts Using the Lightcycler-T(9;22) Quantification Kit
    real-time-quantification 09.05.2000 19:18 Uhr Seite 8 Real-time Quantification of BCR-ABL mRNA Transcripts Using the LightCycler-t(9;22) Quantification Kit Heiko Wittor 1, Hermann Leying 1, Andreas Hochhaus 2, and Rob van Miltenburg 1 1 Roche Molecular Biochemicals, Penzberg, Germany 2 III. Medizinische Universitätsklinik, Fakultät für Klinische Medizin Mannheim der Universität Heidelberg, Mannheim, Germany ly different starting concentrations, so that the initial Introduction target concentration can be determined in a single PCR. The concentration of BCR-ABL transcripts is determi- Literature indicates that in 95 % of all subjects with ned relative to the number of transcripts of a control chronic myeloid leukemia (CML) and in 25-30 % of gene, glucose-6-phosphate dehydrogenase (G6PDH). subjects with acute lymphoblastic leukemia (ALL) a The entire procedure from sample preparation to reciprocal translocation between the long arms of quantitative result is performed in 4.5 hours. chromosome 9 and chromosome 22 [t(9;22)] can be found. This translocation or the resulting fusion pro- duct can be detected by a number of techniques, CLER including fluorescent in situ hybridization, Southern CY blotting, western blotting and reverse transcriptase TThe LightCycler System polymerase chain reaction (RT-PCR). Of these techni- The LightCycler System is based on the amplification LIGHT ques, RT-PCR for the chimeric fusion transcript BCR- of target sequences using alternating heated and ABL has received most attention in relation to the ambient temperature cycles. Samples are contained in detection of minimal residual disease because of its glass capillaries with high surface-to-volume ratio, high sensitivity (1).
    [Show full text]
  • Insulin–Insr Signaling Drives Multipotent Progenitor Differentiation Toward Lymphoid Lineages
    Article Insulin–InsR signaling drives multipotent progenitor differentiation toward lymphoid lineages Pengyan Xia,1* Shuo Wang,1* Ying Du,1 Guanling Huang,1,2 Takashi Satoh,3 Shizuo Akira,3 and Zusen Fan1 1Key Laboratory of Infection and Immunity of CAS, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China 2University of Chinese Academy of Sciences, Beijing 100049, China 3Department of Host Defense, Research Institute for Microbial Diseases (RIMD), Osaka University, Suita, Osaka 565-0871, Japan The lineage commitment of HSCs generates balanced myeloid and lymphoid populations in hematopoiesis. However, the un- derlying mechanisms that control this process remain largely unknown. Here, we show that insulin–insulin receptor (InsR) signaling is required for lineage commitment of multipotent progenitors (MPPs). Deletion of Insr in murine bone marrow causes skewed differentiation of MPPs to myeloid cells. mTOR acts as a downstream effector that modulates MPP differentiation. mTOR activates Stat3 by phosphorylation at serine 727 under insulin stimulation, which binds to the promoter of Ikaros, lead- ing to its transcription priming. Our findings reveal that the insulin–InsR signaling drives MPP differentiation into lymphoid lineages in early lymphopoiesis, which is essential for maintaining a balanced immune system for an individual organism. Hematopoiesis is the process of producing all compo- Insulin, as the primary anabolic hormone, modulates a nents of the blood system from hematopoietic stem cells variety of physiological processes, including growth, differ- (HSCs; Naik et al., 2013; Mendelson and Frenette, 2014; entiation, apoptosis, and synthesis and breakdown of lipid, Walter et al., 2015). HSCs are quiescent, self-renewable pro- protein, and glucose (Samuel and Shulman, 2012).
    [Show full text]
  • PSM, a Mediator of PDGF-BB-, IGF-I-, and Insulin-Stimulated Mitogenesis
    Oncogene (2000) 19, 39 ± 50 ã 2000 Macmillan Publishers Ltd All rights reserved 0950 ± 9232/00 $15.00 www.nature.com/onc PSM, a mediator of PDGF-BB-, IGF-I-, and insulin-stimulated mitogenesis Heimo Riedel*,1,2, Nasim Yousaf1, Yuyuan Zhao4, Heping Dai1,3, Youping Deng1 and Jian Wang1 1Department of Biological Sciences, Wayne State University, Detroit, Michigan, MI 48202, USA; 2Member, Barbara Ann Karmanos Cancer Institute, Detroit, Michigan, MI 48201, USA PSM/SH2-B has been described as a cellular partner of Introduction the FceRI receptor, insulin receptor (IR), insulin-like growth factor-I (IGF-I) receptor (IGF-IR), and nerve The platelet-derived growth factor (PDGF) family growth factor receptor (TrkA). A function has been represents key mitogenic and chemotactic factors that proposed in neuronal dierentiation and development but have been exploited by the Simian sarcoma virus v-sis its role in other signaling pathways is still unclear. To oncogene which results in altered PDGF expression further elucidate the physiologic role of PSM we have (Water®eld et al., 1983). Normal cellular PDGF identi®ed additional mitogenic receptor tyrosine kinases appears in three distinct isoforms as any combination as putative PSM partners including platelet-derived of the PDGF A and B chains which act in paracrine growth factor (PDGF) receptor (PDGFR) beta, hepato- and autocrine mechanisms (Heldin, 1993). PDGF cyte growth factor receptor (Met), and ®broblast growth receptor (PDGFR) signal transduction appears to be factor receptor. We have mapped Y740 as a site of a largely membrane delineated event (Hughes et al., PDGFR beta that is involved in the association with 1996).
    [Show full text]