DOCSLIB.ORG

Targeting the Cancer Kinome Through Polypharmacology

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Targeting the Cancer Kinome Through Polypharmacology PERSPECTIVES components, such as KRAS or PTEN, which THERAPEUTIC RESISTANCE — OPINION enable the tumour to bypass the drug target20,21,30. Targeting the cancer kinome Overcoming these resistance mechanisms will require targeting tumour cells at multiple levels, through either single drugs that bind through polypharmacology to multiple proteins31 or cocktails of highly selective inhibitors32. The challenge for the Zachary A. Knight, Henry Lin and Kevan M. Shokat cancer research community is to learn how to predict the best combinations of targets and Abstract | Kinase inhibitors are the largest class of new cancer drugs. then prioritize those combinations for clini- However, it is already apparent that most tumours can escape from the cal testing. This is a daunting task, because inhibition of any single kinase. If it is necessary to inhibit multiple kinases, the number of possible target combinations how do we choose which ones? In this Opinion article, we discuss some of is almost limitless, but clinical trials are slow the strategies that are currently being used to identify new therapeutic and expensive. combinations of kinase targets. Targeting one kinase with multiple drugs If a tumour depends on the activity of a sin- More than 10,000 patent applications for cancer. Clinical trials show that the most gle kinase, then using multiple drugs to target kinase inhibitors have been filed since 2001 effective kinase inhibitors prolong survival that kinase can be effective. This was first in the United States alone1. This massive by only a few months for these cancers12–17. demonstrated in CML, in which early clinical investment has been fuelled by the realiza- Results have been improved by identifying trials showed that more than 90% of patients tion that kinases are intimately involved in markers for patients that are more likely with chronic phase disease responded to the cancer cell growth, proliferation and sur- to respond to kinase inhibitor therapy — BCR–ABL inhibitor imatinib5 (TABLE 1), but vival. Indeed, kinases and their direct regula- such as EGFR mutations in lung cancer18, that a subset of those patients relapsed while tors are among the most frequently mutated ERBB2 overexpression in breast cancer19, on the drug. Disease progression was associ- oncogenes and tumour suppressors2–4. Well and wild-type KRAS in lung and colorectal ated with the emergence of leukaemic cells known examples include the oncogenic cancer20,21 — but even among these sub- bearing mutations in BCR–ABL that block kinases PIK3CA (the p110α subunit of groups, relapse is inevitable for patients with imatinib binding22, suggesting that drugs PI3K), epidermal growth factor receptor disseminated disease. targeting these BCR–ABL mutants would be (EGFR) and BRAF; the Ras family of onco- Why has clinical progress been so chal- effective. Two second-generation BCR–ABL genes, which activate both PI3Ks and Raf; lenging? One reason is that most tumours inhibitors were developed (dasatinib and and the PTEN tumour suppressor, which can escape from the inhibition of any single nilotinib) that retain activity against most of inhibits PI3K signalling. kinase (FIG. 1). This first became clear when the more than 50 clinically observed BCR– Despite the excitement surrounding these resistance mutations in BCR–ABL were ABL resistance mutations, and these drugs targets, clinical progress has been uneven. discovered in patients with CML who were are highly effective against imatinib-resistant Kinase inhibitors have revolutionized the resistant to imatinib22; similar mutations disease33,34. However, a common BCR–ABL treatment of a select group of diseases, such have now been detected in other kinases fol- mutation (T315I) prevents the binding of as chronic myeloid leukaemia (CML) and lowing treatment with kinase inhibitors23–26. all three drugs, and this has emerged as the gastrointestinal stromal tumours (GIST), Alternatively, tumours can acquire drug default allele for many patients on long- which are driven by a single oncogenic resistance through mechanisms that do not term inhibitor therapy22,35. To address this kinase; for these conditions, kinase inhibi- involve mutation of the target (FIG. 1a). These problem, third-generation drugs have been tors have achieved multi-year increases in mechanisms include the activation of sur- developed that potently inhibit BCR–ABL survival5–7. Smaller but significant responses rogate kinases that substitute for the drug T315I. These agents are effective in preclini- have been observed for some cancers that target27 and the inactivation of phosphatases cal models of drug-resistant CML36–39, and are highly dependent on angiogenesis, and to amplify the residual kinase activity that four such compounds are currently in clini- therefore sensitive to inhibitors of vascular persists during drug treatment28. It is also cal trials. Some patients have now survived endothelial growth factor (VEGF) signalling, clear that many tumours possess intrinsic more than 10 years since starting treatment such as renal cell carcinoma8–11. resistance to kinase inhibitors at the time of by undergoing sequential therapy with three Kinase inhibitors have been least effec- initial therapy (FIG. 1b). This can result from generations of BCR–ABL inhibitors40, prov- tive in treating the types of cancer that have the activation of multiple, redundant kinase ing that it is possible to extend the therapeu- the highest mortality rates, such as lung, signalling pathways29 or the presence of acti- tic response in CML by repeatedly targeting breast, colorectal, pancreatic and prostate vating mutations in downstream pathway the same kinase. 130 | FEBRUARY 2010 | VOLUME 10 www.nature.com/reviews/cancer © 2010 Macmillan Publishers Limited. All rights reserved PERSPECTIVES A similar approach has been used to response. It is unclear how broadly this of an oncogene is much less dramatic than target the ERBB2 receptor tyrosine kinase in model applies, because it is unclear how the response in CML, in which even tran- breast cancer. Trastuzumab is a monoclonal many tumours are truly dependent on a sin- sient inhibition of BCR–ABL irreversibly antibody that binds to the extracellular gle oncogene (a state referred to as ‘oncogene commits cells to apoptosis44 (FIG. 2). In this domain of ERBB2, thereby both inhibiting addiction’ (REF. 41)). The strongest evidence respect, it is worth noting that the term ERBB2 signalling and recruiting immune in favour of this hypothesis is the discovery oncogene addiction gained widespread use cells to the tumour19; however, patients of resistance mutations after kinase inhibitor because it describes a paradox: inhibiting an with metastatic cancer who are treated with treatment in CML, GIST, lung cancer and a oncogene would be predicted to reverse the trastuzumab invariably relapse. The mecha- myeloproliferative disorder known as hyper- gain of function caused by the oncogene, not nism of trastuzumab resistance is not under- eosinophilic syndrome22–26. Such mutations kill all the tumour cells. It is only recently stood, but it is clear that resistant tumours are definitive proof that the mutated kinase that this term has been conflated with the remain dependent on ERBB2 signalling. was required for the survival of that tumour. idea that oncogenes should be expected to This is because patients with breast can- The oncogene addiction model is also sup- be required for tumour survival. For this cer who have progressed on trastuzumab ported by many preclinical studies showing reason, there is clearly a need to identify therapy nonetheless respond to lapatinib14, that tumour cell lines containing an activat- additional vulnerabilities in tumours beyond a small molecule inhibitor of the tyrosine ing mutation or amplification of a kinase can the genes that are directly mutated. kinase domain of ERBB2. Therefore, it is be more sensitive to inhibitors of that kinase possible to induce a second response in these in vitro42–44. Targeting nodes in a signalling network patients by targeting ERBB2 with a drug Conversely, the detection of an oncogenic Three sets of targets collectively account that binds to a different site on the protein. kinase mutation does not guarantee sensitiv- for a large proportion of current efforts in Unlike CML, however, the clinical response ity to the corresponding kinase inhibitor. For kinase inhibitor drug discovery. These are to lapatinib in metastatic breast cancer is example, mutations in PIK3CA or PTEN are the receptor tyrosine kinases (for example, brief, and disease progression typically poor predictors of the sensitivity of tumour EGFR, ERBB2, platelet-derived growth occurs within a few months14. cell lines to PI3K inhibitors31,45. Mutations in factor receptor (PDGFR) and VEGF These examples show that in certain cases KRAS do not, in general, sensitize tumour receptor 2 (VEGFR2)), the kinases in the sequential targeting of a single kinase with cells to inhibitors of Raf or Mek43,46. Indeed, MAPK pathway (for example, BRAF, MEK1 multiple drugs can prolong the therapeutic the response of most tumours to inhibition and MEK2) and the kinases in the PI3K pathway (for example, PIK3CA, Akt and mTOR). These three groups of targets are a Mutation that blocks Upregulation of Downregulation drug binding second kinase of phosphatase mechanistically linked because most recep- tor tyrosine kinases activate the MAPK and PI3K pathways as their primary signalling (FIG. 3) D Drug T315I D D D D function . There is a compelling biological
Load more

Recommended publications
  • Overview and Perspective on RTK Heterointeractions Michael D
    Review Cite This: Chem. Rev. 2019, 119, 5881−5921 pubs.acs.org/CR The RTK Interactome: Overview and Perspective on RTK Heterointeractions Michael D. Paul and Kalina Hristova* Department of Materials Science and Engineering, Institute for NanoBioTechnology, and Program in Molecular Biophysics, Johns Hopkins University, Baltimore, Maryland 21218, United States ABSTRACT: Receptor tyrosine kinases (RTKs) play important roles in cell growth, motility, differentiation, and survival. These single-pass membrane proteins are grouped into subfamilies based on the similarity of their extracellular domains. They are generally thought to be activated by ligand binding, which promotes homodimerization and then autophosphorylation in trans. However, RTK interactions are more complicated, as RTKs can interact in the absence of ligand and heterodimerize within and across subfamilies. 19 Here, we review the known cross-subfamily RTK heterointeractions and their possible biological implications, as well as the methodologies which have been used to study them. Moreover, we demonstrate how thermodynamic models can be used to study RTKs and to explain many of the complicated biological effects which have been described in the literature. Finally, we discuss the concept of the RTK interactome: a putative, extensive network of interactions between the RTKs. This RTK interactome can produce unique signaling outputs; can amplify, inhibit, and modify signaling; and can allow for signaling backups. The existence of the RTK interactome could provide an explanation for the irreproducibility of experimental data from different studies and for the failure of some RTK inhibitors to produce the desired therapeutic effects. We argue that a deeper knowledge of RTK interactome thermodynamics can lead to a better understanding of fundamental RTK signaling processes in health and disease.
    [Show full text]
  • A Molecular Dynamics Simulation Study for Variant Drug Responses Due to FMS-Like Tyrosine Kinase 3 Cite This: RSC Adv.,2017,7, 29871 G697R Mutation†
    RSC Advances View Article Online PAPER View Journal | View Issue A molecular dynamics simulation study for variant drug responses due to FMS-like tyrosine kinase 3 Cite this: RSC Adv.,2017,7, 29871 G697R mutation† Chien-Cheng Lee,a Yu-Chung Chuang,b Yu-Lin Liub and Chia-Ning Yang *b FMS-like tyrosine kinase 3 (FLT3) is an attractive target for acute myeloid leukemia. Recent studies have suggested that the application of small-molecule kinase inhibitors is a promising treatment strategy for patients with primary activating mutations of FLT3; however, the development of secondary mutations, including those of A627T, N676D, F691I, and G697R, that confer acquired resistance to kinase inhibitors has become a severe problem. In this study, we conducted a series of molecular dynamics simulations on PKC412- and sorafenib-bound FLT3 kinases and different apo forms of the FLT3 kinase to explain the minor and severe G697R mutation-induced resistance to sorafenib and PKC412, respectively. Structural analysis on our simulation results revealed that the type II kinase inhibitor sorafenib (IC50 ¼ 9 nM) Creative Commons Attribution 3.0 Unported Licence. assesses its binding site through either the adenine pocket entrance or the back pocket entrance, whereas the type I kinase inhibitor PKC412 (IC50 ¼ 35 nM) intercalates to its binding site from the front pocket entrance. The G697 residue is located at the end of the FLT3 kinase hinge segment and is close to the front and adenine pockets. In G697R mutation where the substituted R697 residue affects both the front and adenine pocket entrances in different manners, sorafenib may approach its binding site Received 11th April 2017 through the back pocket entrance, whereas PKC412 is blocked by the FLT3 kinase.
    [Show full text]
  • Marine Drugs
    marine drugs Article Neuroprotective Effect of Carotenoid-Rich Enteromorpha prolifera Extract via TrkB/Akt Pathway against Oxidative Stress in Hippocampal Neuronal Cells Seung Yeon Baek and Mee Ree Kim * Department of Food and Nutrition, Chungnam National University, Daejeon 34134, Korea; [email protected] * Correspondence: [email protected]; Tel.: +82-42-821-6837; Fax: +82-42-821-8887 Received: 27 June 2020; Accepted: 17 July 2020; Published: 19 July 2020 Abstract: In this study, we found that E. prolifera extract (EAEP) exhibits neuroprotective effects in oxidative stress-induced neuronal cells. EAEP improved cell viability as well as attenuated the formation of intracellular reactive oxygen species (ROS) and apoptotic bodies in glutamate-treated hippocampal neuronal cells (HT-22). Furthermore, EAEP improved the expression of brain-derived neurotrophic factor (BDNF) and antioxidant enzymes such as heme oxygenase-1 (HO-1), NAD(P)H quinine oxidoreductase-1 (NQO-1), and glutamate–cysteine ligase catalytic subunit (GCLC) via the tropomyosin-related kinase receptor B/ protein kinase B (TrkB/Akt) signaling pathway. In contrast, the pre-incubation of K252a, a TrkB inhibitor, or MK-2206, an Akt-selective inhibitor, ameliorated the neuroprotective effects of EAEP in oxidative stress-induced neuronal cells. These results suggest that EAEP protects neuronal cells against oxidative stress-induced apoptosis by upregulating the expression of BDNF and antioxidant enzymes via the activation of the TrkB/Akt pathway. In conclusion, such an effect of EAEP, which is rich in carotenoid-derived compounds, may justify its application as a food supplement in the prevention and treatment of neurodegenerative disorders. Keywords: Enteromorpha prolifera; oxidative stress; apoptosis; BDNF; TrkB/Akt pathway 1.
    [Show full text]
  • Short-Term Rapamycin Persistently Improves Cardiac Function After Cessation of Treatment in Aged Male and Female Mice
    Short-term rapamycin persistently improves cardiac function after cessation of treatment in aged male and female mice. Ellen Quarles A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy University of Washington 2017 Reading Committee: Peter Rabinovitch, Chair Michael MacCoss David Marcinek Program Authorized to Offer Degree: Pathology © Copyright 2017 Ellen Quarles University of Washington Abstract Short-term rapamycin persistently improves cardiac function after cessation of treatment in aged male and female mice. Ellen Quarles Chair of the Supervisory Committee: Peter Rabinovitch, Professor and Vice Chair of Research Department of Pathology Cardiac aging is an intrinsic process that results in impaired cardiac function and dysregulation of cellular and molecular quality control mechanisms. These effects are evident in the decline of diastolic function, increase in left ventricular hypertrophy, metabolic substrate shifts, and alterations to the cardiac proteome. This thesis covers the quality control mechanisms that are associated with cardiac aging, results from an anti-aging intervention in aged mice, and a review of mitochondrial dysfunction in the heart. Chapter one is a review of the quality control mechanisms in aging myocardium. Chapter two consists of the results of several mouse experiments that compare the cardiac function, proteomes, and metabolomes of aged and young controls, along with rapamycin treated aged mice. The novelty of this study comes from the inclusion of a group of animals treated only transiently with the drug, then followed for eight weeks post-drug-removal. This persistence cohort may hold clues to deriving long-lasting benefits of rapamycin with only transient treatment.
    [Show full text]
  • The Role of Neurotrophin Receptors in Female Germ-Cell Survival in Mouse and Human Norah Spears1,*, Michael D
    Research article 5481 The role of neurotrophin receptors in female germ-cell survival in mouse and human Norah Spears1,*, Michael D. Molinek1, Lynne L. L. Robinson2, Norma Fulton2, Helen Cameron1, Kohji Shimoda1, Evelyn E. Telfer3, Richard A. Anderson2 and David J. Price1 1Biomedical Sciences, University of Edinburgh, Hugh Robson Building, George Square, Edinburgh EH8 9XD, UK 2Medical Research Council Human Reproductive Sciences Unit, Centre for Reproductive Biology, The University of Edinburgh Chancellor’s Building, 49 Little France Crescent, Edinburgh EH16 4SA, UK 3Institute of Cellular and Molecular Biology, University of Edinburgh, Darwin Building, Kings Buildings, Edinburgh, UK *Author for correspondence (e-mail: [email protected]) Accepted 4 July 2003 Development 130, 5481-5491 © 2003 The Company of Biologists Ltd doi:10.1242/dev.00707 Summary During mammalian ovary formation, the production of follicle formation in the mouse. In situ hybridisation ovarian follicles is accompanied by an enormous loss of showed that TrkB was expressed primarily in the germ cells germ cells. It is not known how this loss is regulated. We before and after follicle formation. Mouse neonatal and have investigated the role of the Trk tyrosine kinase fetal ovaries and human fetal ovaries were cultured in the receptors, primarily TrkB, in this process. The ovaries of presence of K252a, a potent inhibitor of all Trk receptors. TrkB–/– and TrkC–/– mice with a mixed (129Sv × C57BL/6) In mice, K252a inhibited the survival of germ cells in newly genetic background were examined shortly after birth. formed (primordial) follicles. This effect was rescued by the Around 50% of TrkB–/– mice had grossly abnormal ovaries addition of basic fibroblast growth factor (bFGF) to the that contained greatly reduced numbers of follicles.
    [Show full text]
  • Chemical Biology of Natural Indolocarbazole Products: 30 Years Since the Discovery of Staurosporine
    The Journal of Antibiotics (2009) 62, 17–26 & 2009 Japan Antibiotics Research Association All rights reserved 0021-8820/09 $32.00 www.nature.com/ja REVIEW ARTICLE Chemical biology of natural indolocarbazole products: 30 years since the discovery of staurosporine Hirofumi Nakano and Satoshi O¯ mura Staurosporine was discovered at the Kitasato Institute in 1977 while screening for microbial alkaloids using chemical detection methods. It was during the same era that protein kinase C was discovered and oncogene v-src was shown to have protein kinase activity. Staurosporine was first isolated from a culture of Actinomyces that originated in a soil sample collected in Mizusawa City, Japan. Thereafter, indolocarbazole compounds have been isolated from a variety of organisms. The biosynthesis of staurosporine and related indolocarbazoles was finally elucidated during the past decade through genetic and biochemical studies. Subsequently, several novel indolocarbazoles have been produced using combinatorial biosynthesis. In 1986, 9 years since its discovery, staurosporine and related indolocarbazoles were shown to be nanomolar inhibitors of protein kinases. They can thus be viewed as forerunners of today’s crop of novel anticancer drugs. The finding led many pharmaceutical companies to search for selective protein kinase inhibitors by screening natural products and through chemical synthesis. In the 1990s, imatinib, a Bcr-Abl tyrosine kinase inhibitor, was synthesized and, following human clinical trials for chronic myelogenous leukemia, it was approved for use in the USA in 2001. In 1992, mammalian topoisomerases were shown to be targets for indolocarbazoles. This opened up new possibilities in that indolocarbazole compounds could selectively interact with ATP- binding sites of not only protein kinases but also other proteins that had slight differences in ATP-binding sites.
    [Show full text]
  • Α-Tocopherol at Nanomolar Concentration Protects PC12 Cells from Hydrogen Peroxide-Induced Death and Modulates Protein Kinase Activities
    Int. J. Mol. Sci. 2012, 13, 11543-11568; doi:10.3390/ijms130911543 OPEN ACCESS International Journal of Molecular Sciences ISSN 1422-0067 www.mdpi.com/journal/ijms Article α-Tocopherol at Nanomolar Concentration Protects PC12 Cells from Hydrogen Peroxide-Induced Death and Modulates Protein Kinase Activities Irina O. Zakharova, Tatyana V. Sokolova, Liubov V. Bayunova, Yulia A. Vlasova, Maria P. Rychkova and Natalia F. Avrova * Department of Comparative Neurochemistry, Institute of Evolutionary Physiology and Biochemistry of Russian Academy of Sciences, Thorez avenue, 44, Saint-Petersburg 194223, Russia; E-Mails: [email protected] (I.O.Z.); [email protected] (T.V.S.); [email protected] (L.V.B.); [email protected] (Y.A.V.); [email protected] (M.P.R.) * Author to whom correspondence should be addressed; E-Mail: [email protected]; Tel.: +7-812-552-7901; Fax: +7-812-552-3012. Received: 9 July 2012; in revised form: 23 August 2012 / Accepted: 4 September 2012 / Published: 14 September 2012 Abstract: The aim of this work was to compare protective and anti-apoptotic effects of α-tocopherol at nanomolar and micromolar concentrations against 0.2 mM H2O2-induced toxicity in the PC12 neuronal cell line and to reveal protein kinases that contribute to α-tocopherol protective action. The protection by 100 nM α-tocopherol against H2O2-induced PC12 cell death was pronounced if the time of pre-incubation with α-tocopherol was 3–18 h. For the first time, the protective effect of α-tocopherol was shown to depend on its concentration in the nanomolar range (1 nM < 10 nM < 100 nM), if the pre-incubation time was 18 h.
    [Show full text]
  • Trkb Receptor Signalling: Implications in Neurodegenerative, Psychiatric and Proliferative Disorders
    Int. J. Mol. Sci. 2013, 14, 10122-10142; doi:10.3390/ijms140510122 OPEN ACCESS International Journal of Molecular Sciences ISSN 1422-0067 www.mdpi.com/journal/ijms Review TrkB Receptor Signalling: Implications in Neurodegenerative, Psychiatric and Proliferative Disorders Vivek K. Gupta 1,*, Yuyi You 1, Veer Bala Gupta 2, Alexander Klistorner 1,3 and Stuart L. Graham 1,3 1 Australian School of Advanced Medicine, Macquarie University, F10A, 2 Technology Place, North Ryde, Sydney, NSW 2109, Australia; E-Mails: [email protected] (Y.Y.); [email protected] (A.K.); [email protected] (S.L.G.) 2 Centre of Excellence for Alzheimer’s Disease Research & Care, School of Medical Sciences, Edith Cowan University, Joondalup, WA 6027, Australia; E-Mail: [email protected] 3 Save Sight Institute, Sydney University, Sydney, NSW 2000, Australia * Author to whom correspondence should be addressed; E-Mail: [email protected]; Tel.: +61-2-98-123-537; Fax: +61-2-98-123-600. Received: 27 March 2013; in revised form: 27 April 2013 / Accepted: 28 April 2013 / Published: 13 May 2013 Abstract: The Trk family of receptors play a wide variety of roles in physiological and disease processes in both neuronal and non-neuronal tissues. Amongst these the TrkB receptor in particular has attracted major attention due to its critical role in signalling for brain derived neurotrophic factor (BDNF), neurotrophin-3 (NT3) and neurotrophin-4 (NT4). TrkB signalling is indispensable for the survival, development and synaptic plasticity of several subtypes of neurons in the nervous system. Substantial evidence has emerged over the last decade about the involvement of aberrant TrkB signalling and its compromise in various neuropsychiatric and degenerative conditions.
    [Show full text]
  • Tumor-Associated Lymphatic Endothelial Cells Promote Lymphatic Metastasis by Highly Expressing and Secreting SEMA4C
    Published OnlineFirst July 8, 2016; DOI: 10.1158/1078-0432.CCR-16-0741 Cancer Therapy: Preclinical Clinical Cancer Research Tumor-associated Lymphatic Endothelial Cells Promote Lymphatic Metastasis By Highly Expressing and Secreting SEMA4C Jun-Cheng Wei1, Jie Yang1, Dan Liu1, Ming-Fu Wu1, Long Qiao1, Jun-Nai Wang1, Quan-Fu Ma1, Zhen Zeng1, Shuang-Mei Ye1, En-Song Guo1, Xue-Feng Jiang1, Lan-Ying You1, Ying Chen1, Li Zhou1, Xiao-Yuan Huang1, Tao Zhu1, Li Meng1, Jian-Feng Zhou1, Zuo-Hua Feng2, Ding Ma1, and Qing-Lei Gao1 Abstract Purpose: Lymphatic vessels are mainly regarded as passive associated LECs also produced soluble SEMA4C (sSEMA4C). conduits for the dissemination of cancer cells. In this study, we Increased serum sSEMA4C was detected in patients with breast investigate whether and how the tumor-associated lymphatic cancer and cervical cancer. Patients with metastasis had much vessels may play an active role in tumor metastasis. higher levels of serum sSEMA4C. sSEMA4C promoted lymphan- Experimental Design: In situ laser capture microdissection of giogenesis by activating PlexinB2-ERBB2 signaling in LECs, and lymphatic vessels followed by cDNA microarray analysis was promoted the proliferation and migration of tumor cells by used to determine the expression profiling of lymphatic endo- activating PlexinB2-MET signaling, thus promoting lymphatic thelial cells (LEC). Gene expression levels and activity of signaling metastasis. Although the SEMA4C signaling pathways differ pathways were measured by real-time RT-PCR, ELISA, or immu- between LECs and tumor cells, RHOA activation was necessary noblotting. Lymphangiogenesis was assessed by IHC. Lymph for the effects of SEMA4C in both types of cells.
    [Show full text]
  • Structural and Binding Studies of Human CAMKK2 Kinase Domain Bound to Small Molecule Ligands
    bioRxiv preprint doi: https://doi.org/10.1101/538157; this version posted February 1, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Structural and binding studies of human CAMKK2 kinase domain bound to small molecule ligands Gerson S. Profeta 1,2, Caio V. dos Reis 1,2, Paulo H. C. Godoi 1,2, Angela M. Fala 1,2, Roger Sartori 2, Anita P. T. Salmazo 2, Priscila Z. Ramos 1,2, Katlin B. Massirer 1,2, Jonathan M. Elkins 2,3, David H. Drewry 4, Opher Gileadi 3 & Rafael M. Couñago 1,2 * 1. Centro de Química Medicinal (CQMED), Centro de Biologia Molecular e Engenharia Genética (CBMEG), Universidade Estadual de Campinas (UNICAMP), Campinas, SP, 13083-875, Brazil 2. Structural Genomics Consortium, Departamento de Genética e Evolução, Instituto de Biologia, UNICAMP, Campinas, SP, 13083-886, Brazil 3. Structural Genomics Consortium, University of Oxford, Old Road Campus Research Building, Roosevelt Drive, Oxford, OX3 7DQ, UK 4. Structural Genomics Consortium, UNC Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA * Electronic address: [email protected] bioRxiv preprint doi: https://doi.org/10.1101/538157; this version posted February 1, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license.
    [Show full text]
  • N-Acetyl Serotonin Derivatives As Potent Neuroprotectants for Retinas
    N-acetyl serotonin derivatives as potent neuroprotectants for retinas Jianying Shena, Kanika Ghaib, Pradoldej Sompola, Xia Liua, Xuebing Caob,1, P. Michael Iuvonec, and Keqiang Yea,1 Departments of aPathology and Laboratory Medicine and cOphthalmology and Pharmacology, Emory University School of Medicine; Atlanta, GA 30322; and bDepartment of Neurology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China Edited by Solomon H. Snyder, Johns Hopkins University School of Medicine, Baltimore, MD, and approved January 24, 2012 (received for review November 22, 2011) N-acetylserotonin (NAS) is synthesized from serotonin by arylalkyl- (13, 14). Intraocular injection of NGF, BDNF, and NT-4/5 res- amine N-acetyltransferase (AANAT), which is predominantly ex- cues RGCs after axotomy (15–18). NT-4/5, in combination with pressed in the pineal gland and retina. NAS activates TrkB in a other trophic factors, is involved in the postnatal survival of circadian manner and exhibits antidepressant effects in a TrkB-de- retinal neurons during both development and degeneration (19). pendent manner. It also enhances neurogenesis in hippocampus in Despite the fact that BDNF is not required for RGC survival sleep-deprived mice. Here we report the identification of NAS deriv- during development (20), it promotes survival of RGCs in a va- atives that possess much more robust neurotrophic effects with riety of experimental lesion models both in vitro and in vivo (15, improved pharmacokinetic profiles. The compound N-[2-(5-hydroxy- 21–24). This is consistent with the findings that RGCs express 1H-indol-3-yl)ethyl]-2-oxopiperidine-3-carboxamide (HIOC) selectively the BDNF receptor TrkB (20, 25, 26).
    [Show full text]
  • Tropomyosin-Related Kinase B Signaling Pathway Contributes to The
    Laboratory Investigation (2017) 97, 1332–1342 © 2017 USCAP, Inc All rights reserved 0023-6837/17 Brain-derived neurotrophic factor/tropomyosin-related kinase B signaling pathway contributes to the aggressive behavior of lung squamous cell carcinoma Keigo Ozono1,2, Yoshihiro Ohishi1, Hideya Onishi3, Katsuya Nakamura3,4, Junichi Motoshita5, Masato Kato6, Ryoichi Nakanishi7, Masafumi Nakamura2 and Yoshinao Oda1 The tropomyosin-related kinase (Trk) family consists of TrkA, TrkB, and TrkC, which play essential roles in tumor progression and/or suppression in various cancers. Little is known about the biological significance of the Trk family in human lung squamous cell carcinoma (SCC). Here we investigated the clinical significance of the protein expression of Trk family members in samples from 99 SCC patients, and we explored the relationship between invasion/proliferation activities and Trk expression using lung SCC cell lines to clarify the biological significance of the Trk family in lung SCC. Immunohistochemical high expression of TrkB was significantly correlated with vascular invasion (P = 0.004), lymph node metastasis (Po0.001), and advanced stage (P = 0.0015). The overall survival of the patients with TrkB-high expression was significantly shorter than those with TrkB-low expression (P = 0.0110). TrkA/TrkC expressions were not predictors of poor prognosis. An in vitro assay demonstrated that the inhibition of brain-derived neurotrophic factor (BDNF) (a TrkB ligand) and TrkB by K252a (a Trk inhibitor) or siRNA (BDNF-siRNA, TrkB-siRNA) suppressed the invasion, migration, and proliferative activities of lung SCC cells. The administration of recombinant human BDNF (rhBDNF) enhanced the invasion, migration, and proliferation activities, which were abrogated by K252a.
    [Show full text]