Francisco A Gonzalez-scarano, Professor And Chair
University Of Pennsylvania 3451 Walnut Street Philadelphia, Pa 19104
Grant 5P30AI045008-099001 from National Institute Of Allergy And Infectious Diseases IRG: ZAI1
Abstract: The major goal of the Penn-CFAR Developmental Core is the promotion of AIDS and HIV-related research at the University of Pennsylvania, the Children s Hospital of Philadelphia, and the Wistar Institute by providing a source of funding for pilot projects. The Developmental Core will support short term (1 year) projects in emerging areas of HIV research, including new research related to the recruitment of new investigators, research by established investigators who are turning their attention to AII3S, and research by AIDS/HIV investigators who are undertaking studies that represent a significant departure from their previous work. During the past funding period, we funded a total of 18 projects ranging in scope from molecular analysis of viruses and virus proteins, through the potential development of vaccine candidates, to sociological analysis of the transmission of sexual information in some African societies. A significant proportion of these pilot projects have been converted into fully funded NIH grants. We are proposing to fund 5 projects each year in years 6-10 of this CFAR, adding a new grant category devoted exclusively to interdisciplinary and/or translational research projects.
Keywords: AIDS, biomedical facility, health science research support, human immunodeficiency virus 1
Project start date: 2007-07-01
Project end date: 2009-06-30
Sponsored Links Excellgen http://Excellgen.com
Francisco A Gonzalez-scarano, Professor And Chairman
University Of Pennsylvania 3451 Walnut Street Philadelphia, Pa 19104
Grant 5P30AI045008-089001 from National Institute Of Allergy And Infectious Diseases IRG: ZAI1
Abstract: The major goal of the Penn-CFAR Developmental Core is the promotion of AIDS and HIV-related research at the University of Pennsylvania, the Children s Hospital of Philadelphia, and the Wistar Institute by providing a source of funding for pilot projects. The Developmental Core will support short term (1 year) projects in emerging areas of HIV research, including new research related to the recruitment of new investigators, research by established investigators who are turning their attention to AII3S, and research by AIDS/HIV investigators who are undertaking studies that represent a significant departure from their previous work. During the past funding period, we funded a total of 18 projects ranging in scope from molecular analysis of viruses and virus proteins, through the potential development of vaccine candidates, to sociological analysis of the transmission of sexual information in some African societies. A significant proportion of these pilot projects have been converted into fully funded NIH grants. We are proposing to fund 5 projects each year in years 6-10 of this CFAR, adding a new grant category devoted exclusively to interdisciplinary and/or translational research projects.
Keywords: AIDS, biomedical facility, health science research support, human immunodeficiency virus 1
Grants awarded to Francisco A Gonzalez-scarano
PATHOGENESIS OF PERSISTENT VIRAL INJURY TO THE CNS
Francisco A Gonzalez-scarano, Professor And Chairman
University Of Pennsylvania 3451 Walnut Street Philadelphia, Pa 19104
Grant 5P01NS030606-05 from National Institute Of Neurological Disorders And Stroke IRG: NSPB
Abstract: This proposal brings together a group of investigators with expertise in different viral systems and centers around the common theme of interactions of viruses with the nervous system. The overall goal is a better understanding of the relationships between viral proteins and the determinants of neural cell differentiation, particularly as they relate to viral tropism and viral pathology. The proposal contains 5 major components 4 projects and 1 core. In project I, Dr. Gonzalez-Scarano will look at the interaction of two HIV viral proteins with neural cells and neural cell components; in project II, Dr. Gaulton will study the neuropathology of Gross murine leukemia virus; in project III, Dr. Weiss will study the cytopathology of murine coronaviruses in neural cells, with particular emphasis on the fusion process; in project IV Drs. Eisenberg and Cohen will identify the receptor molecule(s) responsible for Herpes Simplex entry into nervous system cells. Dr. Pleasure will direct a cell culture core that will provide specialized cells and analytical expertise to all of the component projects. The findings from each of these component projects will enhance our understanding of neurologic dysfunction mediated by viruses, and will have applicability to the study of multiple sclerosis, viral encephalitis and AIDS.
Keywords: central nervous system disorder, latent virus infection, nervous system infection, virus infection mechanism
Project start date: 1993-01-01
Project end date: 1998-07-14
5P01NS030606-05 (1997): $883508
5P01NS030606-03 (1995): $790188
5P01NS030606-02 (1994): $755675
1P01NS030606-01A1 (1993): $729556
5P01NS030606-10 (2002): $1106398
5P01NS030606-09 (2001): $1075071
5P01NS030606-08 (2000): $1043808
5P01NS030606-07 (1999): $1013479
2P01NS030606-06A1 (1998): $1002260
CHARACTERIZATION OF THE LA CROSSE VIRUS GLYCOPROTEIN FUSION PEPTIDE
Francisco A Gonzalez-scarano, Professor
University Of Pennsylvania, 3451 Walnut Street, Philadelphia, Pa 19104
Grant 3R01AI074626-02S1 from National Institute Of Allergy And Infectious Diseases
Abstract: This award is issued in response to Notice OD-09-060, Recovery Act Administrative Supplements Providing Summer Research Experiences for Students and Science Educators.La Crosse virus (LACV), a NIAID Category B priority pathogen, is a common cause of pediatric encephalitis and aseptic meningitis in areas of the Midwestern United States where its principal mosquito vector, Ochlerotatus (formerly Aedes) trisariatus, resides. The structural components of the LACV genome (L, M, and S) have essential, well- defined roles in virus pathogenesis. Previous studies from our laboratory using wild-type LACV and TAHV 181/57, a highly neurovirulent strain with low neuroinvasiveness, have mapped the neuroinvasive phenotype to the M segment, which encodes Gn, Gc, and a non-structural protein, NSm. More recently, using recombinant glycoproteins we demonstrated that the region corresponding to the membrane proximal two-thirds of Gc, amino acids 860-1442, is critical in mediating fusion and cell entry. Further computational analysis identified structural similarities between LACV Gc amino acid region 970-1350 and the E1 fusion protein of two alphaviruses Sindbis virus and Semliki Forrest virus (SFV). Collectively, these studies suggested that the LACV Gc, like the alphavirus E1 and the flavivirus E, functions as a type II fusion protein. Within Gc there is a 22 amino acid hydrophobic segment, 1066-1087, that is predicted to correlate structurally with a hydrophobic domain of SFV and Sindbis virus E1. The short sequence is highly conserved within the family Bunyaviridae and features several conserved cysteine residues, as do other type II proteins, such as SFV E1. Based on these features, and in our mutagenesis studies, our working hypothesis is that the LACV Gc (1066-1087) functions as its fusion peptide. In the first specific aim, we will extend these studies by analyzing fusion in mosquito cells, and by identifying peptides and antibodies that further associate this region with fusion and entry. In the second specific aim, we will use a newly developed reverse genetics system to construct LACV mutants incorporating the knowledge gained from the studies on the isolated glycoproteins. These viruses will then allow us to extend our in vitro findings to a mouse model of LACV encephalitis previouly developed by our group (third specific aim). Importantly, as this hydrophobic region is highly conserved among the Bunyaviridae, this proposal will also elucidate mechanisms of virus fusion and entry among other emerging bunyaviruses including the NIAID Category A and C pathogens CCHFV and RVFV and will have significant implications for anti-viral therapy. La Crosse Virus is a common cause of pediatric encephalitis and aseptic meningitis in the Midwestern United States where it principal mosquito vector, Ochlerotatus triseriatus resides. We have identified the fusion peptide for the La Crosse virus glycoprotein Gc. The studies outlined in this proposal will define mechanisms of fusion and entry for La Crosse Virus in both the mammalian and insect host, determine the role of the newly identified fusion domain in the neuropathogenesis of LACV encephalitis, and develop anti-viral therapies (fusion peptide inhibitors and attenuated virus vaccines)
Keywords: ALV related virus; Aedes; Aedes (genus); Africa; Alpha Virus; Alpharetrovirus; Alphavirus; Amino Acids; Antibodies; Arbovirus Encephalitis; Arbovirus, Group B; Arboviruses, Group A; Area; Arthropod-Borne Encephalitis; Aseptic Meningitis; Attenuated; Avian Leukosis-Sarcoma Viruses; Avian Retrovirus; Body Tissues; Bunyamwera virus; Bunyaviridae; Bunyavirus; Categories; Cell fusion; Cells; Central Nervous System; Childhood; Chimera Protein; Chimeric Proteins; Clinical; Collaborations; Computer Analysis; Congo Virus; Congo hemorrhagic fever virus; Crimean-Congo Hemorrhagic Fever Virus; Culicidae; Cysteine; Encephalitis; Encephalitis, Arbovirus; Encephalitis, Epidemic; Encephalitis, Herpes Simplex; Epidemic; Family; Flavivirus; Four Corners Virus; Four Corners hantavirus; Fusion Protein; Gene Products, RNA; Genetic Alteration; Genetic Change; Genetic defect; Genetics-Mutagenesis; Genome; Genus Alpharetrovirus; Glycoproteins; Half-Cystine; Hemorrhagic Fever Virus, Crimean; Hemorrhagic Fever Virus, Crimean-Congo; Herpes encephalitis; Herpetic Encephalitis; Immunity; In Vitro; Incidence; Infection; Inflammation, Brain; Insecta; Insects; Invertebrates, Insects; Knowledge; L-Cysteine; La Crosse virus; LaCrosse Virus; Laboratories; Location; Mammalian Cell; Mammals, Mice; Maps; Mediating; Membrane; Mice; Middle West U.S.; Midwest U.S.; Midwest US; Midwestern United States; Modeling; Molecular Biology, Mutagenesis; Mosquitoes; Muerto Canyon Virus; Murine; Mus; Mutagenesis; Mutation; NIAID; National Institute of Allergy and Infectious Disease; Nervous System, CNS; Neuraxis; Neuropathogenesis; Ochlerotatus; Ochlerotatus (genus); Orthobunyavirus; Peptide antibodies; Peptides; Phenotype; Play; Process; Proteins; RNA; RNA, Non-Polyadenylated; Recombinants; Recurrence; Recurrent; Reporting; Retroviruses, ALV-Related; Ribonucleic Acid; Rift Valley fever virus; Role; Sin Nombre virus; Sindbis Virus; Structural Protein; System; System, LOINC Axis 4; Technology; Tissues; Type C Avian Retroviruses; Vaccines; Viral; Viral Diseases; Viral Encephalitis, Arthropod-Borne; Virus; Virus Diseases; Viruses, General; Work; aminoacid; avian type C retrovirus group; base; computational analysis; design; designing; epizootic; flavivirus glycoprotein E; flaviviruses glycoprotein E; gene product; genome mutation; glycoprotein E, flavivirus; glycoprotein E, flaviviruses; inhibitor; inhibitor/antagonist; member; membrane structure; mouse model; mutant; neurovirulence; neutralizing antibody; pathogen; pediatric; positional cloning; reverse genetics; social role; therapeutic development; vector mosquito; viral infection; virus infection; virus pathogenesis
Project start date: 2009-07-14
Project end date: 2010-09-30
Budget start date: 14-JUL-2009
Budget end date: 30-SEP-2010
PFA/PA: PA-07-070
3R01AI074626-02S1 (2009): $21118
Sponsored Links Excellgen http://Excellgen.com
Francisco A Gonzalez-scarano, Professor
University Of Pennsylvania, 3451 Walnut Street, Philadelphia, Pa 19104
Grant 2T32NS007180-27A1 from National Institute Of Neurological Disorders And Stroke
Abstract: The proposed program will train predoctoral, MD, and PhD postdoctoral investigators in the field of neurovirology. The goal of the program is to develop trainees such that they can develop research programs that can further our understanding of the interaction between viruses and the nervous system, including their potential use as vectors to deliver genes into CNS cells. Generally, the trainees are physicians or veterinarians with prior training in neurology or neuropathology or PhD scientists with prior training in virology or molecular biology or neuroscience. The trainees are given a solid background in virology and through formal coursework introduced to neurological problems where the discipline can play a role. The training program is laboratory based, and includes experience in many of the important methods in virology and molecular biology in the setting of an interactive group. The institution also has an extensive didactic program in neurosciences, virology, microbiology, and gene therapy that can be individually tailored to each trainee in order to supplement the laboratory experience. Each trainee will undertake an independent project that will provide experience in the design and analysis of experiments, and in the presentation and publication of results. Several trainee research seminars (Virology and Neurovirology) provide constant interchange between trainees and trainers. The laboratory work is supplemented by an organized postdoctoral training program that introduces the trainees to important issues that will influence their careers, such as grant writing. It also contains excellent lectures and programs in the mandatory areas such as bioethics. The program is conducted by 11 individual trainers. Each trainee selects one of the laboratories for his/her primary research project, but has free access to other trainers for advice, technical help, and collaboration. An executive committee is in charge of monitoring the trainees´ progress. Each trainer has independent funding, and all have active research programs in neurovirology. In addition, trainees have access to methods and instruments in laboratories of other trainers. Core services are available within the University of Pennsylvania include protein sequencing, peptide and oligonucleotide synthesis
Keywords: Neurovirology; Training
Project start date: 1980-07-01
Project end date: 2014-06-30
Budget start date: 1-JUL-2009
Budget end date: 30-JUN-2010
2T32NS007180-27A1 (2009): $138254
5T32NS007180-26 (2007): $106645
5T32NS007180-25 (2006): $132533
5T32NS007180-24 (2005): $219841
5T32NS007180-23 (2004): $17990
2T32NS007180-22 (2003): $209006
Francisco A Gonzalez-scarano, Professor And Chairman
University Of Pennsylvania 3451 Walnut Street Philadelphia, Pa 19104
Grant 2P30AI045008-069001 from National Institute Of Allergy And Infectious Diseases IRG: ZAI1
Abstract: The major goal of the Penn-CFAR Developmental Core is the promotion of AIDS and HIV-related research at the University of Pennsylvania, the Children s Hospital of Philadelphia, and the Wistar Institute by providing a source of funding for pilot projects. The Developmental Core will support short term (1 year) projects in emerging areas of HIV research, including new research related to the recruitment of new investigators, research by established investigators who are turning their attention to AII3S, and research by AIDS/HIV investigators who are undertaking studies that represent a significant departure from their previous work. During the past funding period, we funded a total of 18 projects ranging in scope from molecular analysis of viruses and virus proteins, through the potential development of vaccine candidates, to sociological analysis of the transmission of sexual information in some African societies. A significant proportion of these pilot projects have been converted into fully funded NIH grants. We are proposing to fund 5 projects each year in years 6-10 of this CFAR, adding a new grant category devoted exclusively to interdisciplinary and/or translational research projects.
Keywords: AIDS, biomedical facility, health science research support, human immunodeficiency virus 1
Project start date: 2004-07-01
Project end date: 2009-06-30
G PROTEIN COUPLED RECEPTORS AND HIV DEMENTIA
Francisco A Gonzalez-scarano, Professor And Chairman
Neurologyuniversity Of Pennsylvania
3451 Walnut Street
philadelphia, Pa 19104
Grant 5R01MH058958-04 from National Institute Of Mental Health IRG: NIVA
Abstract: The identification of members the G-protein coupled, seven-transmembrane domain, receptor (GPCR) family as coreceptors for cellular entry of human immunodeficiency virus (HIV) was a landmark finding that has begun to define at a mechanistic level many of the pathogenetic observations and tropism differences that have been described in association with HIV infection. The CNS contains many GPCRs, including most of the major chemokine receptors that have been implicated in HIV infection as well others that can function as HIV coreceptors in fusion and other assays, but whose function in actual infections is not yet known. Significantly, CCR3 may have a specific role in tropism of viruses for microglial cells (He at al., 1997), and hippocampal neurons express CXCR4 and CCR3. The principal hypotheses in this proposal are that the expression of chemokine receptors and other GPCRs within the CNS plays a major role in the development and pathogenesis of HIV dementia by (1) potentially defining the HIV strains that can establish a chronic CNS infection, (2) serving as a potential route for neuronal pathology mediated by HIV, perhaps by mediating binding by gp120, and, (3) possibly serving as primary receptors for HIV infection of CD4-negative cells. Concomitantly, chemokine expression in the CNS may influence HIV strain variability and neuronal function, since chemokine receptors have been detected in neurons. These hypotheses will be tested with three major specific aims (1) We will determine which chemokine receptors are used by isolates in the CNS, primarily using envelopes cloned directly from infected brains and a combination of assays including cell-to-cell fusion, preparation of pseudotype viruses, and HIV infection of cells transiently transfected with individual chemokine receptors (2) We will determine which HIV coreceptors are expressed in microglia, neurons, and other cells of the CNS using immunohistochemistry, in-situ hybridization, and reverse transcriptase-PCR amplification (RT/PCR) in brain and in cultured CNS cells. We will also determine whether there are differences in the level of expression between brains obtained from HIV-infected individuals with/without HIV encephalitis and individuals with other inflammatory neurological diseases. (3) We will use ELISA and RT-PCR to determine which of the known chemokines are expressed by HIV infected microglia, by other CNS cells, and in HIV infected histological specimens. The results of these studies will further our understanding of the pathophysiology of HIV dementia
Keywords: AIDS dementia complex, G protein, chemokine, cytokine receptor, pathologic process, receptor coupling, receptor expression HIV envelope protein gp120, antireceptor antibody, brain cell, human immunodeficiency virus, neuropathology, virus infection mechanism cell fusion, enzyme linked immunosorbent assay, human tissue, immunocytochemistry, in situ hybridization, polymerase chain reaction, tissue /cell culture, transfection
Project start date: 1998-09-10
Project end date: 2003-06-30
5R01MH058958-04 (2001): $320338
5R01MH058958-03 (2000): $313282
5R01MH058958-02 (1999): $306434
Sponsored Links Excellgen http://Excellgen.com
1R01MH058958-01 (1998): $314687
Short-Term Training For Minority Students
Francisco A Gonzalez-scarano, Professor And Chair
Neurologyuniversity Of Pennsylvania
3451 Walnut Street
philadelphia, Pa 19104
Grant 5T35HL007789-15 from National Heart, Lung, And Blood Institute IRG: ZHL1
Abstract: The goals of the program are to provide qualified undergraduate minority students with the opportunity to conduct biomedical laboratory research and to gain first hand exposure to careers in biomedical sciences through participation in intense 10-12 week summer research experiences at the University of Pennsylvania. The long term aims are two-fold (1) To enhance the total social, personal, and academic perspective of minority students; and (2) To increase the pool of minority students committed to careers in biomedical and biobehavioral sciences, with specific emphasis on heart, lung, and blood diseases, transfusion medicine and sleep disorders. The program is well integrated into all existing educational and research programs of the University of Pennsylvania, and draws on fifteen years of prior experience in the training of over 260 undergraduate minority students. The training faculty consists of established university investigators, including minority faculty, whose research interests focus on cardiovascular, pulmonary, and hematologic questions. The program provides trainees with an in-depth research experience, an opportunity to witness the professional and lifestyle attractions available in the academic community, and information about the training opportunities and financial support for careers in biomedical sciences. Recruitment primarily targets sophomores/juniors at historically black colleges and universities and Hispanic and Native American students. Trainees are selected by the program advisory committee on the basis of academic merit and motivation, and are chosen for laboratory placement by individual mentors based on their interests. The trainees are housed in campus dormitories during their participation in the program. Trainees work on research projects central to the research goals of the laboratory under the direct supervision of the mentor. At the end of the summer, trainees prepare an and an oral presentation of the research for fellow trainees and mentors. Trainees are encouraged to return to Penn for a second summer to deepen their experience. Additional program components include a weekly research seminar, presentations by minority faculty and students, career counseling with directors of graduate research and medical training programs, participation in a national undergraduate research symposium, and peer mentoring for non-academic support
Project start date: 1993-07-01
Project end date: 2009-05-31
5T35HL007789-15 (2007): $52207
5T35HL007789-14 (2006): $188387
5T35HL007789-13 (2005): $188387
5T35HL007789-12 (2004): $156645
2T35HL007789-11 (2003): $183177
Molecular And Cellular Biology Of HIV Encephalopathy
Francisco A Gonzalez-scarano, Professor And Chair
Neurologyuniversity Of Pennsylvania
3451 Walnut Street
philadelphia, Pa 19104
Grant 5P50NS027405-19 from National Institute Of Neurological Disorders And Stroke IRG: ZNS1
Abstract: The CNS is a site of persistent HIV replication and progressive neuropathological damage even in individuals receiving highly active antiretroviral therapy (HAART). CNS HIV reservoirs include both microglia and astrocytes; however, factors controlling HIV replication in these cell types, and the role of microglia and astrocytes in promoting the neurodegeneration characteristic of HIV-associated dementia (HAD) are not fully understood. This represents a large obstacle to the development neuroprotection against HAD. For the next cycle of this program project we propose to further our studies of HAD neuropathogenesis with 4 interactive projects. Dr. F. Gonzalez-Scarano will continue his studies of HIV infection of microglia, with a focus on the role of the association of HIV gag protein with specific exosomal proteins in promoting virus assembly and production in activated macrophages and microglia. Dr. D. Kolson will focus on the mechanisms of astrocyte injury and dysfunction that contribute to neuronal damage, utilizing an in vitro model he recently developed and the technology of in vivo single-cell RNA profiling. Dr. R. Pomerantz, will determine the mechanisms of restriction of HIV replication in astrocytes, based on interactions his group has identified between an astrocytic cellular helicase and the viral regulatory protein Rev. Dr. R. Collman will study the molecular pathways by which HIV triggers macrophage activation through Env/CD4/chemokine receptor interactions, to initiate cascades of cellular activation and mediator secretion that ultimately result in neuronal and glial cell damage. The coordinated studies in this program project will increase our understanding of critical mechanisms of virus persistence and of the pathophysiology of neurodegeneration and provide novel targets for prevention or intervention in HAD
Keywords: AIDS dementia complex, HIV infection, human immunodeficiency virus 1 clinical research
Project start date: 1997-03-01
Project end date: 2010-07-31
5P50NS027405-19 (2008): $1099199
5P01NS027405-18 (2007): $1117170
5P01NS027405-17 (2006): $1133807
2P01NS027405-16A1 (2005): $1176014
Sponsored Links Excellgen http://Excellgen.com
5P01NS027405-13 (2001): $1188269
5P01NS027405-12 (2000): $1186296
2P01NS027405-11 (1999): $1251544
INTERACTIONS OF HIV PROTEINS WITH NEURAL CELLS
Francisco A Gonzalez-scarano, Professor And Chairman
University Of Pennsylvania 3451 Walnut Street Philadelphia, Pa 19104
Grant 5P01NS030606-050001 from National Institute Of Neurological Disorders And Stroke
Abstract: Retroviral infections of the nervous system can result in widespread neurological dysfunction without overt infection of nervous system cells. This has been shown in infections with the Human Immunodeficiency Virus (HIV), where in spite of the relative paucity of infected astro- oligodendroglia or neurons there are significant functional abnormalities in around 20% of cases. To explain this apparent paradox, it has been proposed that modest infection of some central nervous system cells leads to widespread dysfunction by the specific interaction of some viral products with brain cells, either neurons or glia. An alternative hypothesis is that viral proteins produced in either macrophages or microglia-cells shown to be infected in the brain of AIDS patients - may interfere with the function of CNS cells. This proposal will look at the interaction of two HIV proteins, gp120 and tat, with cellular components, particularly at the cell surface, in two specific aims. In aim 1 we will extends experiments that have demonstrated that galactosyl ceramide (GalCer or galactocerebroside), or a closely related molecular plays a significant role in the entry of HIV- 1 into cell lines derived from the human nervous system. In these studies (Harouse et al, 1991; Bhat et al, 1991) we showed that antibodies against GalCer inhibited or decreased infection of two cell lines, U373- MG, derived from glioblastoma, and SK-N-MC, derived from a peripheral neuroblastoma and that gp120 bound GalCer in a specific manner. To extend these findings, we will define the region(s) of gp120 responsible for this interaction using several complementary approaches (i) inhibition of binding by monospecific or monoclonal anti gp120 antibodies (ii) competition for gp120-GalCer binding with peptides (iii) generation of gp120 mutants and (iv) digestion of gp120 with glycosidases. In aim 2 we will study abnormalities induced in cultured neurons by the HIV transactivator, tat, to understand the region of tat responsible for inducing these abnormalities and to map the receptor on brain cells.
Keywords: human immunodeficiency virus, nervous system infection, virus infection mechanism, virus protein, HIV envelope protein gp120, cellular pathology, ceramide, cerebroside, galactose, host organism interaction, latent virus infection, neurophysiology, receptor binding, virus receptor, laboratory rabbit, tissue /cell culture
RESEARCH TRAINING PROGRAM IN DISEASE-ORIENTED NEUROSCIENCE
Francisco A Gonzalez-scarano, Professor
University Of Pennsylvania, 3451 Walnut Street, Philadelphia, Pa 19104
Grant 3R25NS065745-02S1 from National Institute Of Neurological Disorders And Stroke
Abstract: Program The aim of this training program is provide a significant research experience during the neurology residence at the University of Pennsylvania, as a means of accelerating the transition between residency and research career development for our overwhelmingly academically-oriented trainees. The program will be conducted in a large research-oriented institution with a residency program that trains some of the best candidates in the country and that has enough flexibility to encourage a meaningful hiatus from clinical training without compromising the important goals of a neurology residency. Eligible residents will apply to the program through an administrative committee, providing evidence of research aptitude through the development of a research project in conjunction with a research mentor. They will then be able to devote 6-12 months to a project, returning to the residency program at its conclusion. It is expected that the data gathered during this hiatus from clinical training will lead to further research as post-doctoral trainees, and beyond. The training program will be conducted by experienced clinical and laboratory research faculty in various departments with a track record in promoting young careers, including but not limited to Neurology, Neuroscience, Bioengineering, and Medicine. The declining numbers of clinician-scientists is a real threat to the development of new treatments for neurological diseases, as this group populates the ranks of disease-oriented investigators in academia, biotechnology, and pharmaceutical companies. Part of the reason for this decline is the lengthy training process, with the insecurity of future funding. This program will allow residents in neurology to interrupt their clinical training in order to develop clinical or bench-based skills, and to generate sufficient preliminary data to enhance their transition from residency to faculty positions in research-intensive institutions
Keywords: No Project Terms available
Relevance: The declining numbers of clinician-scientists is a real threat to the development of new treatments for neurological diseases, as this group populates the ranks of disease- oriented investigators in academia, biotechnology, and pharmaceutical companies. Part of the reason for this decline is the lengthy training process, with the insecurity of future funding. This program will allow residents in neurology to interrupt their clinical training in order to develop clinical or bench-based skills, and to generate sufficient preliminary data to enhance their transition from residency to faculty positions in research-intensive institutions
Project start date: 2009-03-04
Project end date: 2011-02-28
Budget start date: 1-MAR-2010
Budget end date: 28-FEB-2011
PFA/PA: RFA-NS-09-001
3R25NS065745-02S1 (2010): $151200
Genetics Of Simian Immunodeficiency Virus Encephalopathy
Francisco A Gonzalez-scarano, Professor And Chair
University Of Pennsylvania 3451 Walnut Street Philadelphia, Pa 19104
Grant 5R01MH067734-05 from National Institute Of Mental Health IRG: ZMH1
Abstract: Prior to the widespread use of highly active anti-retroviral therapy (HAART) roughly 20% of HIV-infected patients developed HIV-associated dementia (HAD). HAART has clearly decreased the incidence of HAD, but its prevalence, or that of less severe cognitive disorders, may rise as individuals with HIV infection live longer. Lentiviral encephalitis also occurs consistently in macaques infected with simian immunodeficiency virus (SIV), the most important animal model for HIV. Studies that have analyzed viral genotypes at later points have also established that there is clear genetic divergence between viruses harbored within the brain and the spleen and other peripheral tissue of the same individual, regardless of the portion of the genome that is analyzed. In some instances regional variation within the CNS has also been documented. However it is not clear (1) whether the brain genotypes arise as a result of adaptive selection, decreased immunological constraints due to the brain s privileged position with respect to the immune system, or are due to genetic drift of a compartmentalized population. Alternatively it could be a confluence of all these factors. It is also uncertain (2) whether the variants developing within the brain may be particularly neuropathogenic, and (3) whether the variants within the brain constitute a potential reservoir of latent infection, able to re-seed the systemic circulation after treatment with HAART. We propose to address these questions using simian immunodeficiency virus (SIV) infection of rhesus macaques, a model that recapitulates most of the features of HIV infection including CNS disease. We propose three specific aims. In the first aim, we will amplify and clone env genomic DNA from macaques with SIV encephalitis, compare the genomic DNA with SIV RNA amplified from individual multinucleated giant cells, and assay the envelopes functionally and for their neuropathogenic potential. In the second specific aim, we will analyze blood, cerebrospinal fluid and other tissues in macaques infected with a molecular clone of SIV, to determine the time course of compartmentalization of SIV within the CNS. In the third specific aim we will follow up on previous results indicating that some cells harbor SIV sequences for periods as long as 2 years. These experiments will help clarify the role of genetic sequestration in the development of SIV encephalitis, and by extension, in its human counterpart.
Keywords: AIDS dementia complex, simian immunodeficiency virus, virus genetics, AIDS therapy, apoptosis, central nervous system, encephalitis, gene expression, genotype, lymphocyte, neuropathology, Macaca mulatta, SDS polyacrylamide gel electrophoresis, electron microscopy, immunocytochemistry, polymerase chain reaction
Project start date: 2002-12-01
Project end date: 2008-11-30
5R01MH067734-05 (2007): $398485
5R01MH067734-04 (2006): $406522
5R01MH067734-03 (2005): $494104
5R01MH067734-02 (2004): $482127
Sponsored Links Excellgen http://Excellgen.com
1R01MH067734-01 (2003): $492929
NEUROTROPISM AND MICROGLIAL INVASION
Francisco A Gonzalez-scarano, Professor And Chairman
University Of Pennsylvania 3451 Walnut Street Philadelphia, Pa 19104
Grant 5P01NS027405-120004 from National Institute Of Neurological Disorders And Stroke IRG: ZNS1
Abstract: Microglia are critical to the primary complications of the human immunodeficiency virus (HIV-1) in the central nervous system (CNS), since they are the most commonly infected cell and their infection represents the majority of the viral load in the CNS. This proposal will center on the biology of HIV-1 in microglial in order to develop a better understanding of the role of the virus in the development of this complication, and to identify potential, CNS-specific, treatment strategies in collaboration with the other components of this program. In the first specific aim we will continue our studies on microglial- tropism of HIV-1 isolates using primary isolates from adults and children, and an isolate adapted to microglia bu sequential passage. We will first use a PCR-based assay to analyze the sequential steps of HIV-1 infection in microglia. For those isolates (like the microglia-adapted HIV-1/BORI- 15) which demonstrate a rapid entry phenotype, we will molecularly clones the envelopes, and define the mechanism of enhanced cellular penetration using molecular and biochemical (binding) assays. Where tropism for microglial cells is related to post-entry steps, other portions of the provirus, or the entire proviral genome, will be cloned. We will then determine whether isolates that do not replace to high levels in microglia can nevertheless establish a chronic infection. These isolates will then be used in a SCID-hu model in another Project. In the second specific aim we will determine whether the envelope proteins, and specifically gp120 from isolates with HIV encephalopathy can mediate changes in intracellular free Ca2+ concentrations in monocyte-derived macrophages (MDM), microglia, and other neural cells. Those gp120s that induce intracellular signals will be tested for their ability to mediate apoptosis. In the third specific aim we will determine whether microglial infection by certain HIV isolates results in increased production of chemokines, which could be responsible for increased cellular trafficking into the CNS, and potential amplification of a chronic infection. The result from these experiments will strengthen knowledge about the interactions between HIV-1 and microglial cells.
Keywords: AIDS dementia complex, HIV infection, human immunodeficiency virus 1, microglia, neuropathology, virus genetics, virus replication, HIV envelope protein gp120, antiAIDS agent, calcium, chemokine, gene amplification, intracellular transport, macrophage, molecular cloning, monocyte, neurotropic virus, neutralizing antibody, programmed cell death, viremia, virulence, virus cytopathogenic effect, human tissue, polymerase chain reaction, tissue /cell culture
ROLE OF MICROGLIA IN HIV/DEMENTIA
Francisco A Gonzalez-scarano, Professor And Chairman
University Of Pennsylvania 3451 Walnut Street Philadelphia, Pa 19104
Grant 5R01NS035743-08 from National Institute Of Neurological Disorders And Stroke IRG: ZRG1
Abstract: Microglia are critical the primary complications of the human immunodeficiency virus (HIV-1) in the central nervous system (CNS), since they are the most commonly infected cell and their infection represents the majority of the viral load in the CNS. This proposal will center on the biology of HIV-1 in microglia in order to develop a better understanding of the role of the virus in the development of this complication, and to help identify potential, CNS-specific treatment strategies. In the first specific aim we will continue our studies on microglial- tropism of HIV-1 isolates using primary isolates from adults and children, and an isolated adapted to microglial by sequential passage. We will first use a PCR-based assay to analyze the sequential steps of HIV-1 infection in microglial. For those isolates (like the PCR-based assay to analyze the sequential steps of HIV-1 infection in microglia. For those isolates (like the microglia-adapted HIV-1/BOR-15) which demonstrate a rapid entry phenotype, we will molecularly clone the envelopes, and define the mechanism of enhanced cellular penetration suing molecular and biochemical (binding) assays. Where tropism for microglial cells is related to post-entry steps, other portions of the pro-virus, or the entire proviral genome, will be cloned. We will then determine whether isolates that do not replicate to high levels in microglia can nevertheless establish a chronic infection. In the second specific aim we will determine whether the envelope proteins, and specifically gp120 from isolates with HIV encephalopathy can mediate changes in intracellular free Ca2+ concentrations in monocyte-derived macrophages (MDM), microglia, and neural cells. Those gp120s that induce intracellular signals will be tested for their ability to mediate apoptosis. In the third specific aim we will determine whether microglial infection by certain HIV isolates results in increased production of chemokines, which could be responsible for increased cellular trafficking into the CNS, and potential amplification of a chronic infection. The results from these experiments will strengthen knowledge about the interactions between HIV-1 and microglial cells.
Keywords: AIDS dementia complex, host organism interaction, human immunodeficiency virus 1, microglia, virus infection mechanism, HIV envelope protein gp120, HIV infection, apoptosis, calcium, central nervous system disorder, chemokine, cytokine, macrophage, virulence, virus envelope, clinical research, human tissue, molecular cloning, polymerase chain reaction
Project start date: 1996-12-01
Project end date: 2004-11-30
5R01NS035743-08 (2004): $278700
5R01NS035743-05 (2001): $258327
2R01NS035743-04 (2000): $252684
Francisco A Gonzalez-scarano, Professor And Chairman
Neurologyuniversity Of Pennsylvania
3451 Walnut Street
philadelphia, Pa 19104
Grant 5R01NS035743-03 from National Institute Of Neurological Disorders And Stroke IRG: ARRG
Abstract: Macrophages and microglia are the central cells in the development of HIV dementia (HIVD) macrophages may be responsible for transporting virus into the central nervous system (CNS), and they and microglia are the major cell type harboring HIV sequences. Macrophages and microglia may be responsible for the production of neurotoxins and potentially toxic viral gene products like gp12O and tat, and if neuroglia and neurons are infected to any great extent, microglia are undoubtedly the source of virus. One aspect of neurotropism, the ability to preferentially replicate in microglia, has been identified by studying a large panel of isolates obtained at the time of primary HIV infection. With these, as well as viruses isolated from the brains of individuals with HIVD, we propose to study the microglial-virus interactions important in the development of HIVD in three specific aims. In the first aim we will (i) determine the mechanism of accelerated microglial replication using PCR to delineate the sequential steps of the infectious process and (ii) define whether microglial-tropism is determined at the level of cell entry (iii) determine why microglia are inherently more susceptible than MDM to HIV infection and (iv) explore the effects of the state of activation of microglia, and the influence of cytokines present in brain, like tumor necrosis factor alpha (TNFalpha) and interleukin 1 beta (IL1beta) on viral replication. These experiments will test the hypothesis that microglial tropism is a phenomenon whose mechanism can be clearly defined by analyzing the series of steps involved in viral replication. In the second aim we will define the genetic underpinnings of preferential replication in microglia, and in the third aim we will determine whether neurovirulence is strictly correlated with increased replication by studying the effects of viral infection with primary isolates from blood and brain on microglial secretion of potential neurotoxins like TNFalpha and platelet activating factor (PAF). However, since the critical neurotoxins may not yet have been identified by previous strategies, we will also use differential display, a technique that may detect mRNAs that are either increased or decreased in infected microglia, and has the advantage that it will provide an unbiased look at the system. The results of these studies should help delineate the rational treatment of HIVD
Keywords: AIDS dementia complex, human immunodeficiency virus 1, microglia, virus infection mechanism interleukin 1, macrophage, platelet activating factor, recombinant virus, tumor necrosis factor alpha, virulence human tissue, polymerase chain reaction
Project start date: 1996-12-01
Project end date: 1999-11-30
5R01NS035743-03 (1999): $203560
5R01NS035743-02 (1998): $197631
1R01NS035743-01A1 (1997): $191875
CHARACTERIZATION OF THE LA CROSSE VIRUS GLYCOPROTEIN FUSION PEPTIDE
Francisco A Gonzalez-scarano, Professor
University Of Pennsylvania, 3451 Walnut Street, Philadelphia, Pa 19104
Grant 5R01AI074626-03 from National Institute Of Allergy And Infectious Diseases
Abstract: La Crosse virus (LACV), a NIAID Category B priority pathogen, is a common cause of pediatric encephalitis and aseptic meningitis in areas of the Midwestern United States where its principal mosquito vector, Ochlerotatus (formerly Aedes) trisariatus, resides. The structural components of the LACV genome (L, M, and S) have essential, well- defined roles in virus pathogenesis. Previous studies from our laboratory using wild-type LACV and TAHV 181/57, a highly neurovirulent strain with low neuroinvasiveness, have mapped the neuroinvasive phenotype to the M segment, which encodes Gn, Gc, and a non-structural protein, NSm. More recently, using recombinant glycoproteins we demonstrated that the region corresponding to the membrane proximal two-thirds of Gc, amino acids 860-1442, is critical in mediating fusion and cell entry. Further computational analysis identified structural similarities between LACV Gc amino acid region 970-1350 and the E1 fusion protein of two alphaviruses Sindbis virus and Semliki Forrest virus (SFV). Collectively, these studies suggested that the LACV Gc, like the alphavirus E1 and the flavivirus E, functions as a type II fusion protein. Within Gc there is a 22 amino acid hydrophobic segment, 1066-1087, that is predicted to correlate structurally with a hydrophobic domain of SFV and Sindbis virus E1. The short sequence is highly conserved within the family Bunyaviridae and features several conserved cysteine residues, as do other type II proteins, such as SFV E1. Based on these features, and in our mutagenesis studies, our working hypothesis is that the LACV Gc (1066-1087) functions as its fusion peptide. In the first specific aim, we will extend these studies by analyzing fusion in mosquito cells, and by identifying peptides and antibodies that further associate this region with fusion and entry. In the second specific aim, we will use a newly developed reverse genetics system to construct LACV mutants incorporating the knowledge gained from the studies on the isolated glycoproteins. These viruses will then allow us to extend our in vitro findings to a mouse model of LACV encephalitis previously developed by our group (third specific aim). Importantly, as this hydrophobic region is highly conserved among the Bunyaviridae, this proposal will also elucidate mechanisms of virus fusion and entry among other emerging bunyaviruses including the NIAID Category A and C pathogens CCHFV and RVFV and will have significant implications for anti-viral therapy. PUBLIC HEALTH RELEVANCE La Crosse Virus is a common cause of pediatric encephalitis and aseptic meningitis in the Midwestern United States where it principal mosquito vector, Ochlerotatus triseriatus resides. We have identified the fusion peptide for the La Crosse virus glycoprotein Gc. The studies outlined in this proposal will define mechanisms of fusion and entry for La Crosse Virus in both the mammalian and insect host, determine the role of the newly identified fusion domain in the neuropathogenesis of LACV encephalitis, and develop anti-viral therapies (fusion peptide inhibitors and attenuated virus vaccines)
Keywords: ALV related virus; Aedes; Aedes (genus); Africa; Alpha Virus; Alpharetrovirus; Alphavirus; Amino Acids; Antibodies; Arbovirus Encephalitis; Arbovirus, Group B; Arboviruses, Group A; Area; Arthropod-Borne Encephalitis; Aseptic Meningitis; Attenuated; Avian Leukosis-Sarcoma Viruses; Avian Retrovirus; Body Tissues; Bunyamwera virus; Bunyaviridae; Bunyavirus; Categories; Cell fusion; Cells; Central Nervous System; Childhood; Chimera Protein; Chimeric Proteins; Clinical; Collaborations; Computer Analysis; Congo Virus; Congo hemorrhagic fever virus; Crimean-Congo Hemorrhagic Fever Virus; Culicidae; Cysteine; Encephalitis; Encephalitis, Arbovirus; Encephalitis, Epidemic; Encephalitis, Herpes Simplex; Epidemic; Family; Flavivirus; Four Corners Virus; Four Corners hantavirus; Fusion Protein; Gene Products, RNA; Genetic Alteration; Genetic Change; Genetic defect; Genetics-Mutagenesis; Genome; Genus Alpharetrovirus; Glycoproteins; Half-Cystine; Hemorrhagic Fever Virus, Crimean; Hemorrhagic Fever Virus, Crimean-Congo; Herpes encephalitis; Herpetic Encephalitis; Immunity; In Vitro; Incidence; Infection; Inflammation, Brain; Insecta; Insects; Invertebrates, Insects; Knowledge; L-Cysteine; La Crosse virus; LaCrosse Virus; Laboratories; Location; Mammalian Cell; Mammals, Mice; Maps; Mediating; Membrane; Mice; Middle West U.S.; Midwest U.S.; Midwest US; Midwestern United States; Modeling; Molecular Biology, Mutagenesis; Mosquitoes; Muerto Canyon Virus; Murine; Mus; Mutagenesis; Mutation; NIAID; National Institute of Allergy and Infectious Disease; Nervous System, CNS; Neuraxis; Neuropathogenesis; Ochlerotatus; Ochlerotatus (genus); Orthobunyavirus; Peptide antibodies; Peptides; Phenotype; Play; Process; Proteins; RNA; RNA, Non-Polyadenylated; Recombinants; Recurrence; Recurrent; Reporting; Retroviruses, ALV-Related; Ribonucleic Acid; Rift Valley fever virus; Role; Sin Nombre virus; Sindbis Virus; Structural Protein; System; System, LOINC Axis 4; Technology; Tissues; Type C Avian Retroviruses; Vaccines; Viral; Viral Diseases; Viral Encephalitis, Arthropod-Borne; Virus; Virus Diseases; Viruses, General; Work; aminoacid; avian type C retrovirus group; base; computational analysis; design; designing; epizootic; flavivirus glycoprotein E; flaviviruses glycoprotein E; gene product; genome mutation; inhibitor; inhibitor/antagonist; member; membrane structure; mouse model; mutant; neurovirulence; neutralizing antibody; pathogen; pediatric; positional cloning; reverse genetics; social role; therapeutic development; vector mosquito; viral infection; virus infection; virus pathogenesis
Project start date: 2008-06-01
Project end date: 2013-05-31
Budget start date: 1-JUN-2010
Budget end date: 31-MAY-2011
PFA/PA: PA-07-070
5R01AI074626-03 (2010): $345515
5R01AI074626-02 (2009): $347765
Sponsored Links Excellgen http://Excellgen.com