Steven U. Walkley, DVM - US grants

The neuronal storage disorders result from inherited defects in specific lysosomal hydrolases and are characterized by an array of neurological symptoms including profound mental retardation/dementia, motor system derangements, sensory deficits such as blindness or deafness, and seizures. Such disease most often affect children and are invariably fatal. Although significant progress has been made in understanding the molecular genetics of these diseases and in determining the specific enzymatic defects responsible for individual diseases, little is presently understood as to the ways in which the resulting metabolic derangements generate altered brain function. The discovery that certain neuronal storage diseases are characterized y ectopic growth of dendrites in select neuronal populations, and more recently, that GABAergic neurons are predominately affected by the neuroaxonal dystrophy previously believed to occur non-specifically in many types of neurons, have opened new chapters in the study of these diseases. Fully understanding the causes and consequences of both phenomena will likely offer important insights into the regulation of dendrite growth and maintenance of axonal integrity in normal nervous systems, and will contribute important information about the possible reversibility of disease-induced alterations in brain following metabolic correction of the directed at these cytopathologic events characterizing storage disease. These hypotheses relate to (i) the possible association between ectopic dendritogenesis and mental retardation, (ii) the role of abnormalities in ganglioside metabolism in inducing new dendrite growth, and (iii) the possible association between axonal spheroid formation (neuroaxonal dystrophy) occurring predominately in GABAergic neurons and the motor system derangements and seizure tendencies which characterize many of these disorders. Immunocytochemical procedures at the light and electron microscopic levels will be applied to the study of GABAergic neurons and to the subcellular localization of gangliosides in storage diseases. Anterograde lectin transport studies will be directed at determining sources of synaptic input onto ectopic dendrites. Specific antagonists to NMDA receptors will be used to test the possible relationship between activation of these receptors and regulation of dendrite growth on pyramidal neurons. A key element in most of these studies will be the availability of inherited and induced models of neuronal storage disorders which are near-exact replicas of these diseases in children. The use of swainsonine, a reversible alpha-mannosidase inhibitor, will allow for the induction of a neuronal storage disease, alpha-mannosidosis, which can be metabolically normalized following removal of the inhibitor. Collaborative studies directed at bone marrow transplants in an inherited model with the same disease offer the opportunity for detailed analysis of disease reversibility. Taken as a whole, these studies can be expected to generate important new data on the phenomena of ectopic dendritogenesis and neuroaxonal dystrophy as they occur in storage disorders, and better relate these changes to cellular processes in normal nervous systems.

bioimaging /biomedical imaging; tissue /cell preparation

Batten's disease is an inherited neurological disorder affecting humans and also has been documented in a variety of animal species including dogs and sheep. Individuals with this disorder are characterized by normal appearance at birth, followed by insidiously progressive neurological deterioration including retarded mental development and/or dementia, blindness, movement disorders, and seizures. Death is the in- evitable outcome as no treatment is currently available. Only recently has research progress begun to clarify the metabolic basis for this disease in that massive storage of a single protein (subunit c of mitochondrial ATP-synthase) has been established. This finding suggests that Batten's disease is a proteolipid proteinosis, but the primary enzyme defect awaits determination. Furthermore, changes in brain structure and function set in motion by the primary metabolic defect and that lead to the devastating neurological symptoms also are poorly understood. Our goal in these studies is to use a well documented animal model of Batten's disease, canine neuronal ceroid lipofuscinosis, to explore mechanisms of pathogenesis. We have proposed two alternative hypotheses to explain onset and progression of neuronal dysfunction. Based on the observation that cortical GABAergic neurons contain more mitochondria than other cortical neurons, coupled with identification of a mitochondrial enzyme as the major storage product in this disease, we propose that GABAergic neurons are inherently more susceptible to the primary metabolic defect (Hypothesis I). According to this view, early GABAergic cell loss would be predicted to precede pyramidal cell loss, but the ensuing loss of inhibitory function would later accelerate pyramidal cell dysfunction. Alternatively, brain dysfunction in Batten's disease may be due to the same mechanisms believed critical in neuronal storage diseases known to be caused by lysosomal hydrolase defects (Hypothesis II). In these cases ectopic dendrites and associated synaptic connections, axonal spheroids in GABAergic neurons, and alterations in second messenger systems are believed linked to specific aspects of brain dysfunction. We propose to use state-of-the-art electrophysiologic and morphologic techniques to explore these hypotheses. A greater understanding of the mechanisms of brain dysfunction in Batten's e anticipated to give insight not only into the primary metabolic defect in brain, but also into possible ways to treat and/or ameliorate clinical symptoms.

Neuronal storage diseases are inborn errors of metabolism that result from deficient activity of lysosomal hydrolases. The resulting catabolic deficiency leads to an accumulation of undergraded substrates in the digestive-vacuolar (lysosomal) apparatus of cells, and to an expanding cascade of events that eventually compromises cell function. Although individuals with these diseases often appear normal at birth, neurodegenerative changes inevitable ensue. Psychomotor deficiencies can be severe and may include mental retardation, motor system dysfunction, sensory deficits, and seizures. Although intracellular storage in non-neuropathic forms of lysosomal disorders has been successfully ameliorated by bone marrow transplantation (BMT), the application of BMT to storage diseases with neuronal involvement (e.g, Hurler's disease) has been highly controversial. Working with an inherited model of lysosomal alpha-D-mannosidase deficiency (alpha- mannosidosis), we have unequivocally demonstrated not only that this enzyme increases in activity in the CNS post-BMT, but that intraneuronal storage is reversed and/or prevented. Most importantly, we have used an indigogenic histochemical substrate to demonstrate that acidic alpha- mannosidase is present with neurons and other cells of the CNS. This remarkable finding has established the principle of therapeutic efficacy for BMT in neuronal storage diseases and has led us to evaluate treatment in a different type of storage disorder - GM2 gangliosidosis - using an animal model of BETA-D-N-acetylhexosaminidase deficiency. Our preliminary studies reveal a dramatically different result: In spite of significant elevations of Beta-hexosaminidase activity in brain (30% of normal), substrate reduction was not evident and histochemical staining demonstrated that the enzyme was limited to brain microglia/macrophages. We believe that differences in efficacy in the above models can be exploited in the testing of hypotheses on the mechanism underlying successful treatment. The most commonly stated rationale for use of BMT in children is that donor blood monocytes enter brain, differentiate as microglia, and provide a source of enzyme to enzyme deficient brain cells. Alternative hypotheses include uptake of 'free' enzyme derived from the circulation, and 'metabolic filtration' which depends on substrate diffusion out of diseased cells with uptake and degradation by donor cells. None of these hypotheses is proven and we propose to test them using multidisciplinary in vivo and in vitro studies. Transplants will be carried out in out models at different ages to assess the importance of early treatment, and the dynamics of monocyte invasion of brain in the early weeks post-BMT will be critically examined. Therapeutic effectiveness for these studies will be assessed by clinical, biochemical, histochemical, immunocytochemical and histopathologic criteria. Using cell culture, we will determine whether putative bone marrow-derived cells from normal animals have the capacity to transfer lysosomal enzyme to brain cells from affected animals and whether differential secretion or uptake of alpha-mannosidase and Beta- hexosaminidase occurs. Alternative mechanisms leading to substrate depletion also will be tested. Taken together, these multidisciplinary studies will provide valuable insight into mechanism(s) underlying metabolic correction in neurons following BMT and into pragmatic issues related to BMT as therapy for neuronal storage diseases in children.

Batten's disease is an inherited neurological disorder affecting humans and also has been documented in a variety of animal species including dogs and sheep. Individuals with this disorder are characterized by normal appearance at birth, followed by insidiously progressive neurological deterioration including retarded mental development and/or dementia, blindness, movement disorders, and seizures. Death is the in- evitable outcome as no treatment is currently available. Only recently has research progress begun to clarify the metabolic basis for this disease in that massive storage of a single protein (subunit c of mitochondrial ATP-synthase) has been established. This finding suggests that Batten's disease is a proteolipid proteinosis, but the primary enzyme defect awaits determination. Furthermore, changes in brain structure and function set in motion by the primary metabolic defect and that lead to the devastating neurological symptoms also are poorly understood. Our goal in these studies is to use a well documented animal model of Batten's disease, canine neuronal ceroid lipofuscinosis, to explore mechanisms of pathogenesis. We have proposed two alternative hypotheses to explain onset and progression of neuronal dysfunction. Based on the observation that cortical GABAergic neurons contain more mitochondria than other cortical neurons, coupled with identification of a mitochondrial enzyme as the major storage product in this disease, we propose that GABAergic neurons are inherently more susceptible to the primary metabolic defect (Hypothesis I). According to this view, early GABAergic cell loss would be predicted to precede pyramidal cell loss, but the ensuing loss of inhibitory function would later accelerate pyramidal cell dysfunction. Alternatively, brain dysfunction in Batten's disease may be due to the same mechanisms believed critical in neuronal storage diseases known to be caused by lysosomal hydrolase defects (Hypothesis II). In these cases ectopic dendrites and associated synaptic connections, axonal spheroids in GABAergic neurons, and alterations in second messenger systems are believed linked to specific aspects of brain dysfunction. We propose to use state-of-the-art electrophysiologic and morphologic techniques to explore these hypotheses. A greater understanding of the mechanisms of brain dysfunction in Batten's e anticipated to give insight not only into the primary metabolic defect in brain, but also into possible ways to treat and/or ameliorate clinical symptoms.

DESCRIPTION (Verbatim from the Applicant's Abstract: One of the most profound events in the life of a neuron in the mammalian CNS is the development of the dendritic tree, yet little is understood about the events controlling this process. Under normal circumstances dendrites emerge from neurons during early brain development and form a characteristic dendritic arbor which is maintained throughout the life of the individual. Cortical pyramidal neurons, for example, undergo dendritic differentiation after completing migration to the cortical mantle. A period of explosive sprouting and elaboration of dendritic processes is followed by a period of pruning and shaping. For normal pyramidal neurons there is no evidence that new, primary dendritic sprouting occurs later in development. Yet in one group of rare genetic diseases--Tay-Sachs disease and related neuronal storage disorders--cortical pyramidal neurons do undergo a second phase of dendritogenesis. New dendritic membrane is generated principally at the axon hillock area and in time this membrane is covered with normal appearing dendritic spines and synaptic contacts. These "ectopic" dendrites thus form a secondary basilar dendritic system on affected neurons. In our studies of neuronal storage diseases we have discovered that neurons exhibiting ectopic dendrite growth always have one feature in common: They contain elevated levels of GM2 ganglioside. Furthermore, all evidence suggests that the elevation in this ganglioside precedes the formation of these ectopic dendrites. Armed with this discovery, we have recently explored GM2 expression during normal dendritogenesis in the immature brain. Again, GM2 ganglioside is abundantly expressed at precisely the time when normal dendritic sprouting is occurring, and as dendritic trees mature, GM2 expression decreases. GM2 ganglioside is essentially undetectable in most neurons of the normal, mature cerebral cortex. These findings have led us to hypothesize that GM2 ganglioside is a pivotal modulator of cellular mechanisms controlling dendritic outgrowth in cortical pyramidal neurons. The goal of this research proposal is to rigorously test this hypothesis in the normal, developing cerebral cortex using a combination of in vivo and in vitro studies.

DESCRIPTION (provided by applicant): Lysosomal storage disorders result from a wide spectrum of genetic mutations affecting proteins integral to normal lysosomal function. Defects in no less than 50 proteins have been documented to cause lysosomal dysfunction, and new proteins linked to lysosomal disease continue to be discovered. Lysosomal diseases are known to affect many tissues and organs with most significantly impacting the brain, leading to severe cognitive impairment including dementia, ataxia, tremors and related motor system dysfunctions, blindness, deafness and other sensory impairments, psychotic episodes and seizures. This complex array of clinical features is reflected in a diversity of underlying molecular and cellular abnormalities, including ectopic dendritogenesis, neuroaxonal dystrophy, protein aggregation conditions including tauopathy, neurodegeneration, and so forth. We believe that this diversity of impact characteristic of storage diseases affecting brain is best explained by looking beyond the lysosome to broader abnormalities in endosomal, retrosomal, autophagosomal, and proteasomal systems, what we refer to as the 'Greater Lysosomal System'. Importantly, there is also increasing evidence that lysosomal diseases are not simply states of storage but are also states of deficiency, with failure to salvage degraded substrates from diseased lysosomes leading to shortages of critical precursors for other metabolic processes. Taken together, this new concept of lysosomal disease stresses the importance of viewing the lysosome and its processing streams not simply as a passive digestive process but rather as an integral player in far reaching cellular events, from signal transduction to homeostatic regulation. In order to advance understanding of lysosomal diseases affecting brain and to illuminate the role of the greater lysosomal system as a metabolic regulator in normal neurons, we propose a series of research aims to test hypotheses that attempt to explain the development of two of the most well documented but enigmatic features of many lysosomal diseases - ectopic dendritogenesis and neuroaxonal dystrophy. Delineation of these pathogenic pathways and the underlying role of the lysosomal system we believe will open the door to understanding, and treating, commoner neurodegenerative diseases that share similar features, from Fragile X and other dendritopathies to dementias like Alzheimer's disease.

Lysosomal storage disorders are fatal genetic diseases caused by defects in a wide range of proteins associated with the endosomal-lysosomal system. Niemann-Pick type C (NPC) disease is a cholesterol- glycosphingolipid (GSL) storage disorder caused most commonly by defects in NPC1, a transmembrane protein believed critical In retroehdbcyWtrafficking of substrates from iys6s6mes,'ahd in NPG2, a soluble lysosome protein of unknown function. Absence of either protein causes an essentially identical condition with affected children exhibiting progressive neurological decline beginning at 4-6 years of age and with death occurring in the second decade of life. An important observation in terms of therapy is that affected children most often appear normal at birth and only later, after a threshold of intracellular storage and metabolic disruption has been exceeded, do clinical symptoms develop. This important feature indicates that there isa window of opportunity after birth when therapy aimed at correction of the metabolic defect could potentially rescue cells from their disease fate and thereby ameliorate or prevent brain dysfunction. Therapeutic options for NPC disease, however, are very limited, with enzyme replacement and cell-mediated therapies providing little hope of benefit, particularly for NPC1 deficiency since this protein is not secreted by cells. Even gene therapy will likely only be beneficial to transduced cells again due to the lack of transfer of the NPC1 protein between cells. These clear limitations have driven development of a new therapeutic option - drugs that can limit the build-up of offending substrates in brain and other organs - known as substrate reduction therapy (SRT). An initial approach here was a small molecule inhibitor of GSL synthesis (N-butyldeoxnorjirimycin, Zavesca[unreadable]) which we pioneered as a therapy for NPC disease. A more recent finding suggests that a naturally occurring compound, the cholesterol-derived neurosteroid known as allopregnanolone (ALLO), has a similar ability to limit lysosomal storage in NPC disease. While the mechanism by which ALLO is able to achieve this effect is unknown, recent findings suggest a critical feature is its ability to act as a ligand for the pregnane X receptor (PXR) and thereby to exerttranscriptional control over numerous genes, including those controlling sterol synthesis. The overall goals of our study are to optimize the administration and efficacy of SRT agents using the NPC mouse models, to test an expanded number of candidate PXR-ligand compounds and determine their effects,on cholesterol and GSL accumulation, and to determine whether.the-use of SRT agents in combination will lead to even greater efficacy in delaying and/or preventing clinical deterioration in NPC disease. In addition to testing therapies of direct and practical relevance to NPC-affected children, these studies will also further explore the linkage between storage of GSLs and cholesterol in NPC disease and their relationship to NPC1 and NPC2 protein function.

[unreadable] DESCRIPTION (provided by applicant): Lysosomal disease encompasses nearly 60 different genetic disorders that are broadly classified as sphingolipidoses, mucopolysaccharidoses, neuronal ceroid lipofuscinoses, and others. While individually rare, overall incidence is estimated at 1:7500 live births, making lysosomal disease more frequent than phenylketonuria, one of the most common and well known genetic brain diseases. Progress has been made in understanding pathogenesis and developing therapies for some lysosomal diseases, but others - most notably the glycoproteinoses or glycoprotein storage diseases - have historically received less attention. Included here are a-mannosidosis, [unreadable]-mannosidosis, fucosidosis, aspartylglucosaminuria, Schindler disease, galactosialidosis, sialidosis, and the related diseases, mucolipidoses types II (I-Cell disease), IIIA (Pseudo-Hurler Polydystrophy) and IIIC. Each of these diseases is characterized by defects in lysosomal processing of glycoproteins, oligosaccharides and related compounds, and by severe multi-system disease and premature death. The first international workshop on the glycoproteinoses in 2004 was cosponsored by NINDS and ORD, and organized in concert with a family conference hosted by the International Society for Mannosidosis and Related Diseases (ISMRD). Since this meeting, important developments have occurred for the glycoproteinoses in terms of therapy, gene discovery and mutation analysis, initiation of natural history studies, and development of animal models. The research momentum generated by this first meeting has led to planning for a 2nd workshop spearheaded by the ISMRD and to be held in conjunction with its family conference in Ann Arbor, MI on July 26-28, 2007. International experts in the glycoproteinoses have been invited and will join participating families from around the world representing the full range of glycoprotein storage diseases. The aims of this R13 proposal are (1) to enhance the presence at the scientific sessions of new investigators, junior and minority scientists and clinicians and selected specialists in related disciplines, and (2) to disseminate the workshop proceedings through webcasting. While the glycoproteinoses are the key focus of this meeting, new insights into therapy and pathogenesis can also be anticipated to provide important advances for other lysosomal and genetic brain diseases. Lysosomal disease encompasses nearly 60 different rare disorders that as a group represent one of the most common classes of human genetic disease. Progress has been made in understanding pathogenesis and developing therapies for some lysosomal diseases, but others - most notably the glycoprotein storage diseases - have historically received less attention. The 10 diseases in this group include a-mannosidosis, fucosidosis, galactosialidosis, I-Cell disease and so forth, with each characterized by defects in lysosomal processing of glycoproteins leading to brain dysfunction and premature death. Enhancement of research on the glycoproteinoses could provide important breakthroughs for the understanding and treatment of not only these diseases but for all genetic brain disorders. [unreadable] [unreadable] [unreadable]

DESCRIPTION (provided by applicant): This Gordon Research Conference (GRC) on Lysosomal Disease, with its inaugural meeting scheduled for January, 2011, is one of the newest of over 170 annual conferences organized by the GRC, an organization known world-wide because of the high-quality, cutting-edge nature of its conferences. This new Lysosomal Disease meeting has evolved from the longstanding and highly successful Lysosomes and Endocytosis GRC, which for many years now has focused principally on the burgeoning field of intracellular trafficking and endocytic mechanisms, rather than lysosomal function/disease. Yet Lysosomal Disease represents a group of more than 50 disorders caused by defects in a wide spectrum of proteins, many of which remain poorly understood, and all of which compromise the function of the lysosomal system in critical ways and lead to serious illness and death of affected individuals. This meeting thus offers a critical new venue for addressing the major challenges involving lysosomal diseases, including the need for better understanding of (1) pathogenic mechanisms underlying dysfunction of neurons and brain, as well as other cell types/ organs/systems, (2) specific functions of non-enzymatic, enigmatic proteins causing some of the most difficult to treat lysosomal disorders (NPC1 and NPC2, CLN3, TRPML1, others) and (3) the relationship between specific lysosomal diseases and other chronic neurodegenerative conditions (e.g., Niemann-Pick type C and Alzheimer's, Gaucher and Parkinson's, and so forth). Additionally there is the challenge to develop (4) new and innovative therapies for these disorders coupled with (5) appropriate biomarker identification and validation, with this followed by (6) effective clinical trial development. To address these important topics our goal is to invite 40+ leading internationally known scientists and clinicians working in the field of lysosomal disease to join with approximately another 100 attendees, fully one-third of whom we anticipate will be junior investigators, postdoctoral fellows and graduate students who are at the beginning stages of their careers. The resulting discussion and cross fertilization of ideas and approaches we believe will accelerate not only our understanding of the role of the lysosomal system in health and in disease, but also move us closer to effective therapies for many of these disorders. Based on the large number of supporting emails that we have received from across the globe since the announcement of this new meeting by the GRC, we fully believe that this conference is going to meet a real unmet need for a genuine academic forum for the Lysosomal Disease field. Significance: Lysosomal diseases, while individually rare, together exhibit an incidence of 1:5000 to 7000 live births, making them as a group one of the more common families of genetic disorders. Yet academic meetings focused on the molecular and cellular pathogenesis and treatment of these diseases as a group is uncommon at best. We anticipate that the scientific discussions, research presentations and informal interactions between the participants in this new GRC will significantly contribute to advancing our understanding of these important and understudied disorders, and will also set the stage for development of innovative new therapies. This meeting, furthermore, will also foster the development of the all important next generation of scientists and clinicians focused on these disorders. PUBLIC HEALTH RELEVANCE: The Gordon Research Conference on Lysosomal Disease will bring together major leaders who are contributing to the understanding and treatment of lysosomal diseases, including those involved in basic science disciplines (genetics, biochemistry, cell biology, pathology, development and characterization of animal disease models etc.) as well as in the clinical sciences (pediatric geneticists and neurologists, clinical trial designers, etc.) along with graduate students and junior investigators who represent the future generation of researchers with interest in Lysosomal Disease. The scientific presentations, discussions and workshops during this conference will significantly contribute to advancing our understanding of the pathogenesis of lysosomal diseases, in terms of brain and other organs/systems, and to developing natural history studies and biomarker discovery, as well as new and innovative therapeutic strategies. Importantly, we are confident that the collegial and cooperative atmosphere of the GRC and the sharing of ideas and approaches, the capacity for cross fertilization between scientists and clinicians working on different lysosomal diseases with common challenges, will provide a basis for advancement of understanding and treating Lysosomal Disease in a manner unparalleled by other meetings.

DESCRIPTION (provided by applicant): The overarching goal of the Rose F. Kennedy Intellectual and Developmental Disabilities Research Center (RFK-IDDRC) is to improve the lives of children with intellectual and developmental disabilities through research and clinical outreach. Forty years of distinguished history in basic research and clinical care as one ofthe flagship IDDRCs of NICHD, coupled with important recent faculty recruitments and facility developments at Einstein have, in concert, sown the seeds of a rennaissance in the Kennedy Center IDD program. Critical to this process is a new Kennedy Center leadership team providing a vision of cooperation and collaboration whereby bench scientists and clinicians work intertwined toward the common goal of translating studies in simple systems to clinical insight and therapies with direct relevance to patient health. Four research themes embodying both the basic science strengths at Einstein and critical IDD-related clinical issues inherent in the Bronx and surrounding populations have been identified, as have key scientific cores essential for supporting research advances in these areas. Research themes are focused on the overlapping and ail-important areas of Autism Spectrum Disorders, Neurogenetic and Seizure Disorders, Nutritional and Environmental Determinants of Brain Development, and Deafness and Communication Disorders. Six scientific cores are designed to provide the means for both human and animal phenotyping, neuron and whole brain imaging, cell and tissue manipulation, and genetic analyses, with each centered strategically to provide advancements in bench research and patient care. Partners in this effort include the powerful research engines of Neuroscience and Genetics and other key academic departments at Einstein along with the Kennedy Center's clinical affiliates - the Children's Evaluation and Rehabiltiation Center (CERC) and the Children's Hospital At Montefiore (CHAM). These clinical centers and their contributing clinical departments serve one of the neediest urban populations in the country, where intellectual and developmental disabilities occur in economically deprived, and underserved racially-mixed minority populations. These remarkable research and clincial resources, coupled with a reinvorgorate and inspired leadership and program, provide the basis for a new era for the Rose F. Kennedy Center as it strives toward its goal of improving the lives of children with intellectual and developmental disabilities.

DESCRIPTION (provided by applicant): Niemann-Pick type C (NPC) disease is a cholesterol-glycosphingolipid (GSL) lysosomal storage disorder caused most commonly by defects in NPC1, a transmembrane protein believed critical in retroendocytic trafficking of substrates from lysosomes. Most affected children appear normal at birth, develop progressive neurological disease in their early years and die in their second decade. We have pioneered the development of two compounds for this disorder. The first, N-butyldeoxynojirimycin (NB-DNJ) or miglustat is a documented inhibitor of GSL synthesis, whereas the second, hydroxypropyl ¿-cyclodextrin (HPBCD), is an FDA-approved excipient used for drug solublization. Both compounds are efficacious in delaying onset of neurological disease and prolonging life (by 25% and 100%, respectively) in the mouse model of NPC1 disease. Yet neither drug is understood in terms of the precise mechanism responsible for its effectiveness. For miglustat, evidence for sustained reductions in ganglioside storage following oral administration to Npc1 mice is lacking. Similarly, for HPBCD, while both cholesterol and GSL storage are substantially reduced following treatment in Npc1 mice, the mechanism underlying this benefit is completely unknown, and indeed controversy continues even over its ability to cross the blood brain barrier. This proposal will carry out a series of complementary in vivo and in vitro studies employing current and novel reagents and animal models, and quantitative high-resolution imaging, biochemical and genetic evaluations, each directed at treatment mechanisms for NPC disease. Our first two aims are to precisely define HPBCD's mechanism of action in reducing cholesterol/GSL storage in neurons and to critically re-examine and assess miglustat's ability to reduce GSL synthesis as a basis for its beneficial impact on neuron survival. Our third aim uses an unbiased gene analysis approach to explore the full range of metabolic pathways impacted by each drug. Capitalizing on lessons learned in these aims, new combinatorial treatment strategies will be tested in the fourth aim as a means to substantially improve therapy for children with NPC disease.

01. OBJECTIVES The Translational Neuroimaging (TNI) Core provides IDDRC investigators access to state of the art in vivo multimodality whole organism imaging from mouse to man, with special facilities to accommodate pediatric subjects. In addition to state of the art hardware, software and applications, this Core provides essential collaborative human resources to support the design, implementation, and acquisition and analysis phases of neuroimaging investigation. As such, the TNI core is one arm of a comprehensive effort by multiple IDDRC Cores to develop a broad, deep and accessible repository of human and animal data relating to normal development and disease states. The focal point of this core is the Gruss Magnetic Resonance Research Center (MRRC), which houses 9.4 Tesla small animal and 3.0 Tesla human MRI systems, with extensive support facilities.

We are in the midst of a revolution regarding our evolving understanding of the molecular mechanisms underlying normal embryogenesis and neural development, gene-environmental interactions, associated genetic and epigenetic processes and the pathogenesis, classification and treatment of intellectual and developmental disorders (IDDs). The Tissue Engineering and Cellular Reprogramming Core (TECR) has evolved from a traditional Tissue Culture Core in our previous Kennedy Center grant to now providing an extensive, innovative, interdisciplinary, state-of-the-art array of user services to further accelerate the explosive growth in basic, translational and clinical neuroscience knowledge and scientific applications undertaken by Center investigators as a key component of the new Einstein IDDRC application. The broad scientific and technical expertise of our faculty, staff and specialized consultants and the availability of support services (Table F1) ensure that particularly novel and cross-disciplinary investigations can be fostered and adapted to the increasingly sophisticated needs of IDDRC investigators. These services are designed to facilitate research projects employing a wide variety of experimental approaches ranging from the utilization of cellular and molecular biology techniques, genomic, epigenomic and systems biological analyses and neurophysiological approaches to the study of distinct animal and human cellular and tissue preparations. The Core supplies these services in conjunction with other IDDRC Core Facilities and specialized support services by providing extensive training, oversight, facilities and novel and unique resources to expedite these interdisciplinary goals.

The goal of the Neurogenomics Core is to provide the most advanced genomics and epigenomics assays and analyses to support the neurological and neuroscience research goals of the IDDRC community at the RFK-IDDRC. Modern neuroscience and neurology research has an increasing need for sophisticated genome-wide genetic and epigenetic assays in the quest to understand not only disease states but also the normal functioning of the nervous system. With the goals for the IDDRC encompassing areas such as the study of the pathogenesis of autism, the links between nutrition, obesity and brain development, deafness and communication disorders, and the pathogenesis of neurogenetic disorders, both genetic and epigenetic influences are obvious potential factors that must be considered. A strong cutting-edge Neurogenomics Core that supplies access to the most up-to-date sophisticated genomic technologies in a cost-effective and efficient manner is therefore an essential resource within a Center such as this. We will describe how we have assembled, integrated and extended resources across a system of existing core facilities to serve the specific neurogenetic needs of the IDDRC community at Einstein, leveraging the investment of the IDDRC to provide a powerful integrated readily accessible core facility that allows even the most inexperienced users to perform studies productively and successfully. The resources to be assembled are from multiple existing core facilities at Einstein, all developed and run by the Department of Genetics and the Center for Epigenomics. The leadership for this core is drawn from both of these entities, reflecting their commitment to the success of this core within the IDDRC. The Neurogenomics Core Facility will serve the needs of IDDRC investigators.

The objective of the Human Clinical Phenotyping Core (HCP) is to promote excellence in human phenotyping, with a central mission to facilitate research on intellectual and developmental disabilities (IDD) by a diverse interdisciplinary team of investigators across the Einstein campus. The Core maintains a centralized easily searchable de-identified database of participants for access by Einstein investigators that includes information about data collection from the Neurogenomics and Translational Neuroimaging Cores, and provides continuity to HCP-enrolled participants as they traverse the different Cores and engage in research projects. The HCP is essential to the mission of the RFK-IDDRC to advance diagnosis, prevention, and treatment of children with developmental disabilities, and is positioned to serve as the central hub for a variety of RFK-IDDRC investigators for whom comprehensive phenotyping is key to understanding the implications of their work. It increases access to the populations of interest, increases the cost-effectiveness of research endeavors in well-characterized clinical populations, and facilitates collaboration among researchers with varied and complementary skill-sets and backgrounds.

The goal of the Cell and Molecular Imaging (CMI) Core is to provide RFK-IDDRC investigators with a comprehensive package of expert guidance, training and assistance in optical, electron and related microscopy techniques and image processing, together with access to costly state-of-the-art equipment. It is expected that the vast majority of IDDRC investigators will make use of this Core given its widespread and fundamental applicability to biological studies. This Core, previously known as the Morphology Core, has existed at Einstein for over 20 years, and has grown from two widefield light microscopes and one electron microscope to a compendium of multiple facilities. Its recognized broad utility to investigators also garnered generous financial support from the Department of Neuroscience, both to sustain personnel as well as to provide for purchase of additional equipment during the absence of NICHD-sponsored center funding. This permitted continued Core growth and uninterrupted service to investigators both inside and outside of the Kennedy Center. Core resources include specialized widefield microscopes and electron microscopes that are utilized for a wide array of specimens and purposes by IDDRC investigators. Samples come from a variety of animal species and include cells, tissue sections or slices, organotypic preparations or whole organisms. Specific microscopy goals for IDDRC investigators are diverse and include studies focused on areas such as cell organization or cytoarchitecture, intracellular organelle distribution, and changes in pathologic markers. Live cell or tissue imaging studies are also carried out, including tracer studies for analysis of organelle transport and turnover. All studies benefit from expert advice and assistance from experienced Core personnel followed by comprehensive image acquisition and analysis. Utility of the Core is further extended by its provision of equipment for high-resolution laser-assisted dissection for gene expression and proteomic studies on tissue and cell preparations, and biolistic technology for facile delivery of naked DNA constructs. The goal is not only to facilitate high-end equipment use, but also to educate and provide highly personalized advice and support to investigators on all relevant methods from initial specimen preparation to final packaging of data for publication or presentation.

G1. OBJECTIVES The overarching mission of the Animal Behavior (AB) Core is to assist investigators seeking to discover behavioral, physiological and metabolic phenotypes in diverse rodent models of intellectual and developmental disabilities. To achieve this mission, the Core performs studies in mice and rats to identify the functional alterations resulting from genetic, developmental or environmental manipulations that may impair neural and behavioral development. These include changes in developmental milestones, sensorimotor function, cognitive function, affective and social behaviors, feeding and activity patterns, body composition and/or, energy expenditure. Through collaborative efforts with the Neurogenomics (NGEN), Cellular and Molecular Imaging (CMI), Tissue Engineering and Reprogramming(TECR), Human Clinical Phenotyping (HCP) and Translational Neuroimaging (TNI) Cores, the consequences of defined genetic or physiological alterations in mice and rats are thoroughly characterized to determine their impact in the context of the measures most relevant and translatable to the human disease phenotype. The role of candidate molecules in relevant tissues, such as neurons, glia and skeletal muscle can be elucidated by thorough and definitive experimentation and screening in mouse and rat models. To enhance research capabilities specific to IDD-related projects, the IDDRC leadership has leveraged the resources of two existing Einstein Shared Resources to form the AB Core. These are the Rodent Behavioral Evaluation Core established by the Department of Neuroscience and headed by Dr. Gulinello, and the Animal Physiology Core developed by the Diabetes Research and Training Center and headed by Dr. Schwartz. The AB Core is designed to satisfy the diverse needs of all IDDRC investigators using awake, unrestrained rodents in their studies by providing state-of-the-art assessments of developmental cognitive and sensorimotor function, of affective, social and motivated behavior, and of whole body and brain metabolism. The structure of the core reflects the fact that advances in understanding the neurobiology of IDD increasingly require not only classical measures of cognitive, motivated and sensorimotor functions, but must also include assessments of nutritional, metabolic and feeding-related behaviors, that together determine the functional profile of the subject and interact with the gentoype and pharmacological interventions. By combining existing Core capabilities and experienced faculty from the Departments of Neuroscience and Medicine, we have established an Animal Behavior Core uniquely suited to plan, perform and evaluate coordinated behavioral and metabolic assessments in developing and adult rodents.

A2. JUSTIFICATION Dynamic Leadership: The primary function of the Administrative Core (ADM) is to provide strong consistent scientific leadership and to enunciate and execute a clear vision for the scientific direction of the Rose F. Kennedy IDDRC at Einstein. This will be achieved through active maintenance and constant re-evaluation of the six scientific Cores that provide the fundamental infrastructure of the center. The leadership will ensure prioritization of access to these Cores for IDDRC investigators, will consistently monitor Core utilization and cost-effectiveness, and ensure Core optimization and modernization through a system of continued evaluation and quality-control. The leadership will promote the activities and successes of the center and its investigators through dissemination of IDDRC work via our website, internal and external publications and dissemination through national media outlets. The leadership will coordinate and focus efforts around four identified thematic areas of IDD research strength. They will organize and manage a vibrant colloquium and mini-workshop series to promote investigator collaborations and community outreach. The leadership will administer a pilot grant program that will draw new investigators into the field of IDD research and provide seed funds for new innovative work from our established investigators. The Director, Dr. Walkley is an internationally recognized authority in IDD research with proven leadership skills and the requisite passion and determination to relentlessly advance and evolve the agenda of the IDDRC. He will take primary responsibility for executing this vision. He will consult with his associate director. Dr. Foxe, and leverage the collective expertise of his five key advisory committees to set a strong successful and vibrant agenda for the RFK-IDDRC. He will advocate at the highest levels of the college administration to guarantee the prominence of IDD research efforts at Einstein.

DESCRIPTION (provided by applicant): Following from its highly successful inaugural meeting at the Hotel Galvez in Galveston, Texas, in 2011, this second meeting of the Gordon Research Conference (GRC) on Lysosomal Disease will take place at the Renaissance Tuscany Il Ciocco Resort in Lucca (Barga), Italy in April, 2013. The Lysosomal Disease GRC is one of the newest of over 170 annual conferences organized by the GRC, an organization known world-wide for its high-quality, cutting-edge academic conferences. The Lysosomal Disease GRC evolved from the longstanding and highly successful Lysosomes and Endocytosis GRC, which for many years has focused principally on the burgeoning field of intracellular trafficking and endocytic mechanisms, rather than lysosomal function/disease. Yet Lysosomal Disease represents a group of more than 50 disorders caused by inherited defects in a wide spectrum of proteins, many of which remain poorly understood. These disorders affect many organ systems, most notably brain, leading to chronic illness and death of affected individuals. The Lysosomal Disease GRC offers a critical venue for addressing the major challenges involving lysosomal diseases, including the need for better understanding of (1) pathogenic mechanisms underlying the dysfunction of neurons and brain, as well as other cell types/organs/systems, (2) specific functions of non-enzymatic, enigmatic proteins (NPC1 and NPC2, CLN3, TRPML1, others) causing some of the most difficult to treat lysosomal disorders and (3) the relationship between specific lysosomal diseases and other chronic neurodegenerative conditions (e.g., Niemann-Pick type C and Alzheimer's, Gaucher and Parkinson's, and so forth). Additionally there is the challenge to develop (4) new and innovative therapies for these disorders coupled with (5) appropriate biomarker identification and validation, with this followed by (6) effective clinical trial development. To address these important topics we have invited 41 leading and internationally known scientists and clinicians working in the field of lysosomal disease to join with approximately another 160 attendees, fully one-third of whom we anticipate will be junior investigators, postdoctoral fellows and graduate students who are at the early stages of their careers. Importantly, the success of the 2011 meeting led to the GRC Organization selecting the Lysosomal Disease meeting as a host for a Gordon Research Seminar (GRS), a special meeting organized and run by graduate students and early postdoctoral fellows. The presence of the GRS insures that the best and the brightest of the next generation of lysosomal disease researchers will be part of the GRC as well. The discussion and cross fertilization of ideas and approaches occurring as part of the GRC and GRS meetings we believe will accelerate not only our understanding of the role of the lysosomal system in health and in disease, but also move us closer to effective therapies for many of these disorders.

? DESCRIPTION (provided by applicant): The Lysosomal Disease GRC had its first meeting at the Hotel Galvez in Galveston, Texas in 2011, and its second at the Renaissance Tuscany Il Ciocco Resort in Lucca (Barga), Italy in 2013. We are one of the newest of over 170 annual conferences organized by the GRC, an organization known world-wide for its high-quality, cutting-edge academic conferences, and indeed are the only new GRC approved as a repeat conference in their 2013 review. Lysosomal Disease represents a group of more than 50 disorders caused by inherited defects in a wide spectrum of proteins, many of which remain poorly understood. These disorders affect many organ systems, most notably brain, leading to chronic illness and death of affected individuals. The 2015 Lysosomal Disease GRC will again offer a critical venue for addressing major challenges involving lysosomal diseases, including the need for better understanding of (1) the role of the greater lysosomal system in maintaining cell homeostasis in health and disease, (2) lysosomal disease pathogenic mechanisms underlying the dysfunction of neurons and brain, as well as other cell types/organs/systems, (3) specific functions of lysosomal and non-lysosomal proteins, defects in which cause lysosomal disease, (4) the relationship between lysosomal dysfunction and other chronic neurodegenerative conditions (e.g., Alzheimer's and Parkinson's), and (5) the extent to which heterozygosity in autosomal and X-linked lysosomal diseases can lead to cellular and clinical disease. Additionally there is the challenge (6) through rigorous preclinical studies to develop new and innovative therapies for these disorders coupled with (7) initiating and completing effective clinical trials. To address these important topics we have to date invited, as speakers and discussants, 27 leading and internationally known scientists and clinicians working in the field of lysosomal disease. Each has accepted our invitation and will join what we anticipate to be an additional 170 attendees; fully one-third of whom we anticipate will be junior investigators, postdoctoral fellows and graduate students who are at the early stages of their careers. Importantly, the success of our 2011 meeting led to the GRC Organization selecting the Lysosomal Disease meeting as a host for a Gordon Research Seminar (GRS), a special meeting organized and run by graduate students and early postdoctoral fellows. Our 2015 meeting has again been selected to host a GRS. The presence of the GRS insures that the very best and the brightest of the next generation of lysosomal disease researchers will be part of the GRC as well. The discussion and cross fertilization of ideas and approaches occurring as part of the GRC and GRS meetings we believe will accelerate not only our understanding of the role of the lysosomal system in health and in disease, but also move us closer to effective therapies for many of these disorders.

? DESCRIPTION (provided by applicant): Niemann Pick C disease is a rare, neurodegenerative, lipid storage disorder. Approximately 95% of the disease is caused by mutations in NPC1, a late endosomal membrane protein that functions in export of lipoprotein-derived cholesterol. The most prevalent NPC1 mutation, I1061T, produces a protein that is misfolded and rapidly degraded. Histone deacetylase inhibitors (HDACi) recently have been shown to reduce the accumulation of cholesterol and other lipids found in patient cells harboring the NPC1I1061T and other mutations. This beneficial effect is associated with decreased endoplasmic reticulum-associated degradation and enhanced delivery of the mutant NPC1 proteins to late endosomes and lysosomes. With the recent generation in our laboratory of a humanized mouse model in which the I1061T mutation knocked into the murine NPC1 locus, it is possible to examine the effect of HDACi on NPC1 stability in vivo. We hypothesize that treatment with an HDACi in the NPC1I1061T knockin model of NPC1 disease will increase levels of the mutant NPC1I1061T protein, slowing progression of neurodegeneration and prolonging survival. The therapeutic potential of HDACi for treatment of NPC1 disease is being explored in through a collaboration involving pharmaceutical partners and an HDACi collaborative involving investigators from NIH (NICHD/NCATS), Weill Cornell Medical College, University of Notre Dame, Albert Einstein College of Medicine, and Washington University, along with the Ara Parseghian Medical Research Foundation. The goals of this proposal are to identify orally-available, CNS-penetrant HDAC-selective compounds using cell-based screens; to evaluate in vivo in the NPC1I1061T knockin model candidate HDACi compounds; and to develop effective therapeutic regimens for testing of the HDACi in clinical trials. The proposed in vivo studies further will provide valuable data for initial dosing protocols and biomarker monitoring in future human trials.

? DESCRIPTION (provided by applicant): Christianson syndrome (CS) is a recently discovered X-linked neurodevelopmental disorder caused by deleterious mutations in SLC9A6 which encodes the sodium-hydrogen exchanger known as NHE6. This protein, along with 8 other NHE proteins, are members of the family of solute carriers found localized to different membranes in cells where they are thought to influence the pH of luminal areas. NHE6 is associated with early endosomes and recycling endosomes and recent studies of a knockout mouse model have shown defects in growth factor signaling, neurotransmitter receptor cycling as well as in lysosomal function. Patients with CS are clinically recognized by features of intellectual disability, ataxia, epilepsy, minimal verbal status, postnatal microcephaly, and cerebellar degeneration. Autistic features have also been ascribed to some patients. Prior to identification of the CS gene, the presence of a `happy demeanor' typically led to a diagnosis of Angelman syndrome, and thus was referred to as X-linked Angelman-like syndrome. Importantly, males with CS described to date have epilepsy, with seizure types including: infantile spasms, tonic seizures, tonic-clonic seizures, myoclonic seizures, drop seizures, and episodes described as staring spells. Clinical involvement in female carriers of CS has been less closely analyzed to date. While CS is a rare condition, Morrow and colleagues have identified 21 affected families and 26 individuals, which include families and patients from the United States, Canada, and Europe. A Christianson Syndrome Association (CSA) was formed in 2011 by a family in Houston, Texas, and a similar organization has now been established in Canada. The CSA held an initial meeting at Brown University in 2013 (hosted by Dr. Eric Morrow) that brought together families with interested clinicians and scientists. The CSA strongly supports the inclusion of an international scientific conference in conjunction with its 2015 family meeting and the purpose of this proposal is to secure funding to facilitate this event. Planned for this meeting are presentations from a diverse array of well-known speakers and CS experts - national and international, including leaders in the field of neurodevelopmental and seizure disorders. We also anticipate the creation of impactful opportunities for junior investigators, including women and minorities, to participate in scientific exchange and to meet CS patients and their families. Key outcomes expected from this meeting include: (i) networking and establishment of collaborative research and clinical outreach programs; (ii) generation of new ideas on the pathogenesis and possible treatment of CS, including new avenues of research and collaborative grants; (iii) expansion of the CS research and clinical community, including the introduction of junior scientists and clinicians to the importance of studying CS and other related rare diseases; and (iv) establishment of an international CS research and clinical network to foster fully collaborative, multi-laboratory basic research and to encourage initiation of a patien registry and natural history study in order to advance patient care and treatment.

    ABSTRACT The primary function of the Administrative Core (Core A, ADM) is to provide strong, consistent scientific leadership and to enunciate and execute a clear vision for the scientific direction of the Rose F. Kennedy IDDRC at Einstein/Montefiore. This is achieved through active supervision and evaluation of the 4 scientific Cores that provide the fundamental infrastructure of the Center, which are coupled to, and tightly integrated with, our Research Project focused on intellectual disability in 22q11.2 Deletion Syndrome. ADM Core leadership ensures prioritization of access to these scientific Cores for IDDRC investigators and consistently monitors Core utilization, quality control and cost effectiveness. They also ensure scientific Core optimization and modernization through regular monitoring as well as a rigorously pursued annual survey. The leadership promotes the activities and successes of the Center and its investigators through dissemination of IDDRC work via our website, internal and external publications including a biannual newsletter and through national media outlets. ADM personnel organize the annual Isabelle Rapin Conference on Communication Disorders, as well as specialized mini-workshops to promote investigator collaborations and community outreach. ADM is also an ongoing sponsor of IDD-related ?interest groups? on various topics ranging from Rett syndrome to dendrito- genesis, as well as IDD-trainee luncheons for pre- and postdoctoral and clinical fellows. The ADM leadership also oversees a College-funded pilot grant program that draws new investigators into the field of IDD research and provides seed funds for innovative work related to IDD. The RFK Director and Associate Director (Drs. Walkley and Molholm, respectively) work with their Executive Council to lead the Center and rely on other standing committees (Internal and External Advisory Committees and a Parent Advisory Committee) for strategic planning and feedback. They are also active members of the national IDDRC network. The ADM Core is also the major outreach arm that acts to connect basic science labs at Einstein with IDD-focused clinics, including those at the Children's Hospital at Montefiore (CHAM) and the Children's Evaluation and Rehabilitation Center (CERC) at Einstein. Through this committed leadership and vigorous advocacy on behalf of IDD research, the ADM Core aggressively promotes and nourishes excellence in IDD-related basic, clinical and translational research at Einstein/Montefiore.

  PROJECT SUMMARY (OVERALL) The overarching purpose of the Rose F. Kennedy Intellectual and Developmental Disabilities Research Center (RFK IDDRC) is to improve the lives of children with intellectual and developmental disabilities (IDD). Fifty years of distinguished progress in basic, translational and clinical research as one of NICHD's flagship IDDRCs, coupled with important recent faculty recruitments and an historic merger between the Albert Einstein College of Medicine and its University-Affiliated Hospital, Montefiore Medical Center, offer a solid platform for continuing excellence in our commitment to IDD research. The Center's 4 highly integrated and complemen- tary scientific Cores consist of: 1) our clinical translational core known as the Human Clinical Phenotyping (Core B, HCP), which serves to facilitate both access to and characterization of participants for IDD relevant research; 2) a Neurogenomics facility (Core C, NGEN) that provides cutting edge epigenetic and genomic processing and analyses on both human and animal tissues; 3) a Neural Cell Engineering and Imaging facility (Core D, NCEI) that provides state-of-the-art approaches to brain cell manipulation and visual-ization; and 4) an Animal Phenotyping facility (Core E, AP) for evaluation of animal behavior, metabolism, imaging and brain function in a manner with strong parallels to approaches taken in patients accessed through HCP. Each of our scientific Cores is carefully overseen and monitored by the Administrative Core (Core A, ADM), which also serves as the head ganglion of the entire IDDRC in its outreach programs to the Einstein/ Montefiore community as well as nationally. Each scientific Core is tightly connected to our selected Research Project which brings together a multidisciplinary team of investigators focused on intellectual disability in 22q11.2 Deletion Syndrome (22q11.2DS). A central function of the RFK IDDRC is to promote the substantive links between Einstein research laboratories and clinics at the Children's Evaluation and Rehabilitation Center (CERC) Children's and the Hospital at Montefiore (CHAM). Together, this set of Cores and our research project form a dynamic network of IDD-focused programs and practices that interlink 19 different academic departments and 20-plus IDD-relevant clinics at Einstein and Montefiore. The latter include 22q11.2DS, Rett and Williams syndromes, tuberous sclerosis, neurofibromatosis, West syndrome and infantile spasms, autism spectrum disorders, and a wide range of lysosomal diseases. New initiatives being set in motion involve outcome assessments of premature birth in terms of IDD and newborn screening (NBS) and critical genotype/ phenotype issues in conditions of high relevance to such NBS programs in New York State (Krabbe disease and adrenoleukodystrophy). Strong outreach programs driven by our ADM core help spread the word about IDD research. Such efforts over the past 5 years have positioned the RFK IDDRC in a substantive leadership role in coupling research and clinical strategic goals across the newly merged Einstein-Montefiore community, and will play an essential role in promotion of new clinical-research partnerships going forward.

Lysosomal diseases represent a group of nearly 60 monogenic human disorders caused by defects in proteins involved in normal functioning of the lysosomal system. Most severely impact the brain, cause progressive neurological deterioration over years to decades, and are fatal. Pathogenic cascades caused by lysosomal dysfunction are remarkably complex and involve diverse and unusual events ranging from the blockage of autophagy to the growth of bizarre and unique (to lysosomal diseases) ?ectopic? dendrites on cortical pyramidal neurons. To provide a conceptual framework for understanding this complexity we developed in 2009 the concept of a ?Greater Lysosomal System? which put the lysosome at center stage in the cell's recycling process, receiving ?streams? of different metabolites from both endosomal and autophagosomal pathways. We also emphasized ?egress? of catabolic products from lysosomes since lack of such salvage would be anticipated to result in deficient precursors for metabolic pathways and possible up-regulation of synthesis or induction of autophagy to overcome such deficiency. Importantly, recent discoveries give credence to this concept ? most notably that a master regulator of cell metabolism, the mammalian target of rapamycin (mTOR, specifically mTORC1), is anchored at the surface of lysosomes. Here, among a myriad of functions, it controls the translocation of the MITF family of transcription factors (e.g., TFEB, TFE3) which themselves regulate hundreds of genes involved in autophagy and lysosomal biogenesis. Thus much evidence now supports the idea of the lysosome as the cell's ?nutrient sensor?, allowing for orchestration of cell growth programs during periods of high nutrient availability and facilitating autophagy during nutrient starvation. We believe this is the most important window yet discovered through which to investigate the basis for the complexity of pathogenic mechanisms in lysosomal diseases. A central goal of the current proposal is therefore to analyze mTOR function across a carefully selected but diverse group of lysosomal diseases and to do so in concert with our earlier and ongoing studies focused on the heterogeneity of lysosomal storage, the dysregulation of autophagy and p62 aggregation, and the unique growth of new, primary dendrites on cortical pyramidal neurons undergoing lysosomal storage of gangliosides. Thus we propose three highly interlinked specific aims: The first to further characterize lysosomal storage heterogeneity as well as p62 aggregation and its relationship to lysosomes; the second to investigate the impact of lysosomal storage on mTORC1 pathway hypo- and hyperactivation and the consequences of each; and the third to determine the association between altered mTOR activation and changes in dendritic complexity, including so-called ?ectopic dendritogenesis?.

This application is for a new T32 institutional training program to support postdoctoral research in intellectual and developmental disabilities (IDDs). The Rose F. Kennedy Center and its program of IDD research has for 50 years been in the forefront of discoveries in brain development, function and disease. Today, we are in an era of unprecedented discovery of new genes and gene variants, as well as environmental and related conditions, linked to IDD. At the same time, we are armed with remarkable advances in modern research tools in neuroscience, genetics and in cell and molecular biology. Our goal here is to create a vibrant new training program that successfully brings together talented postdoctoral fellows working with world-class investigators and their laboratories to develop the next generation of highly motivated and successful IDD-focused research scientists. The four broad objectives of this program are (1) to provide in-depth research experiences and didactic training in biomedical science for highly talented postdoctoral fellows in a manner that shapes their lifetime career objectives and at the same time fosters discovery and advancement of our understanding of, and ability to treat, specific types of IDDs; (2) to build effective cross-discipline communication between basic research laboratories and IDD-focused clinics, sensitizing our postdoctoral trainees to the perspective of IDD-focused clinicians and vice versa; (3) to extend this network of cooperation between trainees and their laboratories, and clinicians and their clinics, to encompass whenever possible the parents/care providers and IDD-affected patients; and (4) to provide career guidance and professional development to our trainees, with this effort coupled to our goal to recruit and train members of underrepresented minority groups. To accomplish these important goals, we have assem- bled a team of 36 highly talented primary trainers selected from our 100-member Rose F. Kennedy IDDRC membership. These individuals come from 15 basic science and clinical departments and span a host of research approaches to IDD ? from human investigations including iPSC studies, to mouse, fish, fly and worm models of IDDs. In addition, we have developed a structured training program designed to broaden the trainees IDD-focus well beyond the immediate lab experience. We have also established a plan of mentoring committees to guide our trainees along with evaluation processes allowing us to continually monitor and modify our program to ensure we are on track to our stated goals. Finally, we have engaged our clinical partners working with patients with IDDs and established a strong role for them in the training program. This program brings our institution?s considerable depth, rigorous scientific approach, and tradition of innovation to bear on the important study of the cause, treatment and prevention of intellectual disability.

PROJECT SUMMARY/ABSTRACT (Core A: ADMINISTRATIVE ? ADM) The primary functions of the Administrative Core (Core A, ADM) are to provide strong, consistent scientific leadership and to enunciate and execute a clear vision for the scientific direction of the Rose F. Kennedy IDDRC at Einstein/Montefiore. This is achieved through active supervision and evaluation of the 4 scientific Cores that provide the fundamental infrastructure of the Center, and which are coupled to, and tightly integrated with, our Research Project focused on intellectual disability in KDM5C disease. ADM Core directors (Drs. Walkley and Molholm) work tirelessly to build an inclusive and vital community at Einstein/Montefiore focused on IDD research. They act to ensure prioritization of access by IDDRC investigators to these scientific Cores and consistently monitor Core utilization, quality control and cost effectiveness. They also ensure scientific Core optimization and modernization through regular monitoring as well as a rigorously pursued annual survey. The leadership promotes the activities and successes of the Center and its investigators through dissemination of IDDRC work via our website, internal and external publications including an annual newsletter and national media outlets, and through active participation in the National IDDRC Network. ADM personnel organize annually the Isabelle Rapin Conference on Communication Disorders and International Rare Disease Day, as well as specialized mini-workshops to promote investigator collaborations and community outreach. The ADM Core is also the major outreach arm that acts to connect basic science labs at Einstein with IDD-focused clinics, including those at the Children?s Hospital at Montefiore (CHAM) and the Children?s Evaluation and Rehabilitation Center (CERC) at Einstein. This is accomplished by various means, including our College-funded pilot grant program that draws new investigators into the field of IDD research and provides seed funds for innovative work related to IDD, as well as our innovative and highly successful program in precision medicine knows as Operation IDD Gene Team. These efforts are tightly coupled to our newly funded IDD T32 postdoctoral training grant which itself is integrated into ongoing efforts to foster IDD involvement and training of Einstein medical and graduate students. The RFK Directors work with their Executive Committee to lead the Center and rely on other standing committees (Internal and External Advisory Committees and Family Advisory Committee) for strategic planning and feedback. Representation of URMs in training, faculty recruitment and patient enrollment are strongly supported here in our Bronx-based IDDRC. Through committed leadership and vigorous advocacy on behalf of IDD research, the ADM Core aggressively promotes excellence in IDD-related basic, clinical and translational research at Einstein/Montefiore.

PROJECT SUMMARY/ABSTRACT (OVERALL) The overarching purpose of the Rose F. Kennedy Intellectual and Developmental Disabilities Research Center (RFK IDDRC) is to improve the lives of children with intellectual and developmental disabilities (IDDs). Fifty- plus years of distinguished progress in basic, translational and clinical research as one of NICHD?s flagship IDDRCs, coupled with important recent faculty recruitments and an historic merger between the Albert Einstein College of Medicine and its University-Affiliated Hospital, Montefiore Medical Center, offer a solid platform for continuing excellence in our commitment to IDD research. The Center?s 4 highly integrated scientific Cores consist of: 1) our clinical translational core known as the Human Clinical Phenotyping (Core B, HCP), which serves to facilitate both access to and characterization of participants for IDD relevant research; 2) a Neurogenomics facility (Core C, NGEN) that provides cutting edge epigenetic and genomic processing and analyses on both human and animal tissues; 3) a Neural Cell Engineering and Imaging facility (Core D, NCEI) that provides state-of-the-art approaches to brain cell manipulation and visualization; and 4) an Animal Phenotyping facility (Core E, AP) for evaluation of animal behavior, metabolism and imaging in a manner with strong parallels to approaches taken in patients accessed through HCP. Each of our scientific Cores is carefully overseen and monitored by the Administrative Core (Core A, ADM), which also serves as the head ganglion of the entire IDDRC in its substantial outreach programs to Einstein/Montefiore, the Bronx community as well as nationally. Each scientific Core has an essential connection to our signature Research Project which brings together a multidisciplinary team of investigators focused on mechanisms of IDD in children with mutations in the transcriptional regulator lysine demethylase 5c gene, KDM5C. A central function of the RFK IDDRC is to promote the substantive links between Einstein research laboratories and clinics at the Children's Evaluation and Rehabilitation Center (CERC) and the Children?s Hospital At Montefiore (CHAM). Together, our Cores and research project form a dynamic network ? a community ? of IDD-focused programs and practices that interlink 18 different academic departments, >100 IDDRC members and 20-plus IDD-relevant clinics at Einstein and Montefiore. The latter include 22q11.2DS, Rett and Williams syndromes, tuberous sclerosis, neurofibromatosis, West syndrome and infantile spasms, autism spectrum disorders, and a wide range of neurometabolic disorders. New initiatives moving forward at this time include our unique precision medicine/community outreach program we call Operation IDD Gene Team, our goal to fortify ties with our Einstein clinical partner (CERC and its affiliated UCEDD and LEND programs) under the umbrella of the Rose F. Kennedy Center and a heightened focus on training through our newly established T32 for IDD postdoctoral fellows.