Molecular and Cellular Biology; RNA; Virology
My research is dedicated to the folding of biological macromolecules such as ribonucleic acids and proteins into higher-order structures and to the role their conformation plays in the way they exert their function within the cell. In particular, my research group studies RNA structural switches involved in the replication of RNA viruses and subviral RNA pathogens. We also study RNA and protein structures that contribute to the regulation of gene expression in other microbial systems through specific RNA-RNA and RNA-protein interactions. Plus-strand RNA viruses are responsible for many diseases in humans, animals and plants. Our efforts are focused on an early step in the viral life cycle within the host cell, the recruitment of the viral RNA genome into a replication complex with viral and cellular proteins. We use yeast as a model host to express two RNA replicons, turnip crinkle virus associated with satellite RNA, by itself or in the presence of viral replication proteins, and potato spindle tuber viroid RNA. Satellites and viroids are subviral RNAs that do not encode their own proteins; they rely entirely on factors provided by the associated helper virus or the host cell. The smaller size and simpler organization of their genome makes them convenient model systems to investigate the role of RNA structure in recognition by viral and host proteins, structural changes involved in these interactions, molecular evolution and intracellular transport. Our goal is to develop a fully controlled replication system where every component is tractable and tunable using tools from genetics, biochemistry, cellular and molecular biology. With this system, we will be able to screen for RNA replication inhibitors and develop RNA vectors with novel functions. I enjoy teaching and mentoring students from a variety of disciplines in the laboratory as well as in the classroom. I believe that meaningful faculty/student interaction is mutually beneficial: it helps students grow into well-rounded citizen-scientists, researchers or health care professionals, and it helps me become a better educator. In my research group, I deeply value and strive to foster diversity. I believe a diverse team creates a more energizing and successful research environment--one where everyone learns from one another and the range of backgrounds and perspectives adds up to a rich learning environment that is much more than the sum of its parts. I am the course director for, and teach in Microbiology for Allied Health Professionals. On the graduate level, I direct the master’s program and teach in the core course of virology.
The human immunodeficiency virus (HIV) is now considered a chronic disease in the developed world. In underdeveloped areas where access to antiretroviral therapy is limited, however, it remains a devastating disease contributing to grave socioeconomic problems. The goal of my research is to expand our knowledge of pathogen interactions with cellular membranes by developing a detailed understanding of the mechanism of HIV entry and by studying co-infection of HIV with the pathogenic fungus Cryptococcus neoformans in human macrophages. The first step of HIV infection is HIV entry when the envelope protein complex on the surface of the virus comes into contact with the cellular receptors, glycoprotein CD4 and coreceptor, and mediates merging of the viral and cellular membranes leading to delivery of the viral genetic material. Mechanistic studies help to inform the development of inhibitors to HIV entry that will be beneficial on both therapeutic and prophylactic levels. The envelope protein complex is the machinery that gets the virus into the cell; as such, it is also a prime target for the development of vaccines. HIV/AIDS often kills by priming the host for opportunistic infections. Cryptococcal meningitis is one of the leading killers of AIDS patients. The human macrophage is the cell type tasked with ingesting and clearing microbes. In my lab, we are working to define the role of the human macrophage in the copathogenesis of the opportunistic fungus Cryptococcus neoformans and HIV during AIDS progression. The mechanisms of host-microbe interactions also serve as templates for the design of novel drug regimens, including immunotherapy. We have recently utilized our extensive experience in the study of how HIV enters the cell to begin studies in Ebolavirus entry which has a similar mechanism. We are developing inhibitors to the process of Ebolavirus entry and using developments in inhibition to study the mechanism of attachment and membrane fusion into multiple cell types. It is my objective throughout my career to provide vital basic research in virology and cell biology in order to advance medical treatment and prevention. As an academic researcher, I put a strong emphasis on the training and mentoring of young scientists in my lab, and I participate in the T35 training grant from the National Institutes of Health that UB and Roswell Park Cancer Institute jointly secured. I train master’s and PhD students as well as postdoctoral fellows in the departments of Microbiology and Immunology and Biochemistry. I also mentor undergraduates in research projects; these students may come to me independently or through UB’s Center for Undergraduate Research and Creative Activities (CURCA) in which I am active. I direct undergraduate studies for my department, and I am the course director for Biomedical Microbiology, my department’s large undergraduate basic science course.
Anatomic Pathology; Blood Banking/Transfusion Medicine; Clinical Pathology; Cytopathology; Hematology - Clinical Pathology; Immunopathology; Surgical Pathology; Toxicology; Transfusion Medicine; Bioinformatics; Microbiology; Virology
I serve the Department of Pathology and Anatomic Sciences as a general pathologist in anatomic and clinical pathology. My primary areas for service work include surgical pathology and cytopathology as an attending pathologist rotating among the Kaleida hospital sites and clinical pathology activities in clinical chemistry, transfusion medicine, microbiology and hematology. I serve as the laboratory medical director for the clinical laboratories at the Women and Children‘s Hospital of Buffalo and the Center for Laboratory Medicine, Williamsville (Flint). I also provide more specialized medical support for the forensic toxicology laboratory at the Women and Children‘s Hospital, the fetal defect screening program at the Center for Laboratory Medicine in Williamsville, the Virology laboratory at Women and Children‘s Hospital and the Therapeutic Plasmapheresis program at the Buffalo General Medical Center. I have developed an interest in Clinical Informatics and regularly employ those skills to retrieve and analyze data from Kaleida and elsewhere to support clinical decision making, research activities, EHR development and business development. Within the department, I am the pathologist overseeing the Transfusion Service across Kaleida and also provide pathology direction to the Kaleida Clinical Chemistry and Microbiology programs. In addition, I support the leadership of Kaleida in their Utilization Program, the Gainsharing Program, Peer Review and as Chair of the Site specific Transfusion Committees. Since January 2013 I have also served as the laboratory director of the Erie County Public Health Laboratory. Previously I have served as the laboratory director at the Center for Laboratory Medicine, Amherst (Suburban), Buffalo General Hospital and as an assistant laboratory director at Gates Circle. Each of these positions has been valuable to me in learning how different groups work together and how different groups of clinicians see and set expectations for a pathology department. Outside of Kaleida, I serve the region as representative to the Erie County Medical Society Legislative Committee and the Economic Affairs Committee. I have also served as president of the Western New York Society of Pathologists (1999-2000) and as Delegate to the College of American Pathologists House of Delegates (2005 - present). The overall theme of these activities is to leverage the skills cultivated by any practicing pathologist to recognize patterns. Those patterns recognized are then directed to purposes that can be quite diverse, ranging from diagnosis to data integrity. Data retrieved from multiple sources are used to provide an unbiased review for departmental and hospital leaders to troubleshoot, drive test menus or to review patterns of practice. Good data can drive good decisions, but only to the degree that the data can be recognized and understood. My professional time is divided in four parts, with anatomic pathology service work comprising about one quarter of my time, clinical pathology service work a second quarter, administrative activities a third quarter and clinical informatics the last quarter (plus or minus 5%), but with the added bonus that on any one day, these duties can shift dramatically to address the needs of the department and hospital. One of the most rewarding parts of my career has been the opportunity do all of these to the best of my ability and to support the efforts of the excellent professionals around me. The variety of responsibilities I have translate into a job that is never dull. I have used my own situation as a model for the pathology residents I train to provide a live demonstration that the field of Pathology is big enough to have something of interest for any interested person.
Infectious Disease; Microbiology; Molecular and Cellular Biology; Molecular genetics; DNA Replication, Recombination and Repair; Virology; Genome Integrity
The major focus of my laboratory is in understanding the molecular machines that make up the DNA replication forks of the small human DNA viruses, polyoma- and papillomaviruses. Papillomaviruses and polyomaviruses are human pathogens; human papillomavirus (HPV) results in a vast number of human cancers, and the human polyomaviruses JC and BK cause serious disease and death in immunocompromised patients. Both viral systems provide important models for the study of human DNA replication mechanisms and have allowed for vital insights into eukaryotic DNA replication. The study of polyomavirus DNA replication led to the first identification of many cellular DNA replication complexes and processes; papillomavirus has provided the best structures and models to date of replicative hexameric DNA helicases and how they function. I typically train undergraduate, master’s and doctoral students and postdoctoral scholars, assistant research professors and laboratory technicians. My laboratory focuses on two primary areas. One is elucidating the dynamic protein-protein interactions that allow the series of enzymes required to replicate DNA to act in concert and in the correct sequence required to duplicate the genome. My laboratory has been at the forefront of identifying the interactions between the one critical HPV DNA replication protein, the origin-binding DNA helicase, E1, and cellular DNA replication proteins. Understanding these interactions and the roles they play in the HPV DNA replication process has helped our understanding of, and continues to lead to information that tells us more about how both viral and eukaryotic DNA replication forks function. In addition, as we identify protein-protein interactions between HPV E1 and cellular factors that are essential for HPV DNA synthesis, we will uncover potential targets for development of broad-range HPV antivirals that could act to block HPV replication. We recently obtained a large multilaboratory NIH research grant to investigate just this possibility for the interaction between HPV E1 and the human DNA replication protein, Topoisomerase I. The second primary area of investigation is elucidating how the cellular DNA damage response (DDR) pathways inhibit DNA replication when cells are subjected to DNA damage. For many years, the DDR field focused on the effects of DDR on the cell cycle kinases as the only method by which DNA replication was arrested. In the mid- to late-2000s, researchers recognized that in mammalian cells there is also a substantial (tenfold) inhibition of elongation of DNA replication following DDR. The mechanisms for this inhibition are unknown. Using both in vitro and cell-based simian virus 40 (SV40) DNA replication systems, we have shown that SV40 DNA replication is also shut down in response to DDR kinase pathways and that this is not based on cell cycle kinase action. Therefore, SV40 provides a useful model system for determining how elongation of DNA replication is inhibited by DDR. Furthermore, we have shown that in contrast HPV DNA replication does not respond to DDR, providing us an important control DNA replication system for these studies. (The lack of DDR arrest of HPV DNA replication likely explains why HPV integrates so readily into host cell chromosomes−an important step for HPV-induced carcinogenesis). Our studies on the DDR effect on polyoma and papilloma virus DNA replication will lead to insights into the effect of DDR on cellular DNA replication as well as an understanding of how HPV integrates into host cell chromosomes causing HPV-induced cancers.
Gene therapy; Genomics and proteomics; Immunology; Infectious Disease; Neurobiology; Neuropharmacology; Viral Pathogenesis; Virology
As a postdoctoral fellowship in the Division of Allergy, Immunology & Rheumatology at University at Buffalo I received a NIDA funded National Research Service Award (NRSA) F32 to study the mechanisms of cocaine-induced HIV-1 infection in astrocytes. This was a two year fellowship award ($99,224). I received several Young Investigator Travel Awards to attend and present my research at national conferences including the Society for NeuroImmune Pharmacology, the College on Problems of Drug Dependence and the International Society for NeuroVirology. I was the first to demonstrate that cocaine enhances the replication of HIV in astrocytes, specialized glial cells in the central nervous system. During this time I was first author on 3 publications and contributed as a co-author on 6 publications in internationally recognized, peer reviewed journals including the Journal of Immunology, Brain Research and Biochimica et Biophysica Acta. As a Research Assistant Professor in the Division of Immunology I was funded through a NIDA Mentored Research Scientist Development Award (K01) award to investigate targeted nanoparticles for gene silencing in the context of HIV and drug abuse. This K01, was a five year award, $785000 that allowed for advanced training in nanotechnology and immunology. I applied this new expertise in nanotechnology to the development of innovative methods to control HIV-1 infections, particularly those associated with methamphetamine abuse. I was an invited panel speaker at the International Symposium on NeuroVirology and the American College of Neuropsychopharmacology. During this time, I published approximately 30 peer-reviewed publications in internationally recognized, peer-reviewed journals, including journals such as the Journal of Immunology, Brain Research, and the Journal of Pharmacology Experimental Therapeutics. Six as first author, 1 as senior author and 23 as a co-author. Presently, I am a Associate Professor and Proposal Development Officer in the Department of Medicine at University at Buffalo where I continue to develop my research in drug delivery methods. I am currently investigating exosomes as potential delivery vehicles. Exosomes are one of several types of membrane vesicles known to be secreted by cells including microvesicles, apoptotic bodies, or exosome-like vesicles. Exosomes, unlike synthetic nanoparticles, are released from host cells and have the potential to be novel nanoparticle therapeutic carriers I have recently been invited to be a panel speaker at the American Society of Nanomedicine and the American Society of Gene & Cell Therapy conferences. I have been a principal investigator and co-instigator on NIH funded projects studying multimodal nanoparticles for targeted drug delivery and immunotherapy in Tuberculosis and HIV and a co-investigator on a NYS Empire Clinical Research Investigator Program (ECRIP) to develop a Center for Nanomedicine at UB and Kaleida Health. I have had over eight years of NIH supported funding.
Structural Biology; X-ray Crystallography; Bioinformatics; Genomics and proteomics; Infectious Disease; Microbial Pathogenesis; Molecular and Cellular Biology; Protein Function and Structure; Proteins and metalloenzymes; Virology
The overarching goal of the Umland Lab is to use structural biology combined with biochemical, molecular biology, and genetics to explore important elements of infectious disease. The objective is to both extend the fundamental understanding of how microbial pathogens interact with their respective hosts and to identify new antimicrobial targets and new antimicrobial therapeutics. Two major projects on this theme are on going within the lab. In the first, unrecognized and underexploited potential antimicrobial targets within multi-, extreme, and pan-drug resistant gram-negative bacilli (GNB) are being identified and then characterized using the phenotype of in vivo essentiality. That is, our interest is in genes and their corresponding gene products that are essential for bacterial growth and survival during infection of a host (i.e., in vivo) rather than only essential under ideal laboratory growth conditions (e.g., rich laboratory media, absence of immune responses, etc.). The class of genes that are in vivo essential but not in vitro essential has largely been neglected as antimicrobial targets, and so represents a rich set for expanding target space in the urgent race to develop new antimicrobials. The second project is focused upon identifying and characterizing virus protein - host protein interactions. Viruses encode a highly limited set of functionality, and therefore rely on subverting cellular machinery. This high jacking of cellular functions for the benefit of the virus often involved virus-host protein-protein interactions (PPIs). Study of these virus-host PPIs reveals both the mechanisms by which viruses co-opt cellular functions and potential new antiviral targets recalcitrant to the development of drug resistance. An additional rationale for studying virus-host PPIs is to understand virus evolution with respect to PPI involvement in virulence, pathogenesis, and host tropism. In conjunction with both of these projects, the Umland Lab is using structurally enabled fragment-based lead discovery (FBLD) methods to identify small molecules with potential to be developed into antimicrobial therapeutics.