Biochemical investigation of NUBPL disease mechanisms and therapy candidates

Awardee: Eiko Ogiso

Institution: Children's Hospital of Philadelphia

Grant Amount: $120,465.00

Funding Period: February 1, 2023 - January 31, 2024

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Summary:

The goal of this project is to unveil NUBPL disease mechanisms and accelerate identification of therapeutic candidates that improves health in human NUBPL disease patients. Toward this goal, we utilize three NUBPL-/- genetic disease models, C. elegans (worm, invertebrate), D. rerio (zebrafish, vertebrate), and human patient cells. First, we will investigate therapeutic efficacy of top drug candidates recently identified in other complex I disease worms as well as nutrient and signaling pathway modulators (including vitamins and dietary supplements), in our NUBPL-/- zebrafish and human patient fibroblasts. Second, we will expand our focus on iron regulation and metabolism including response to iron therapy, which might also be involved in NUBPL disease pathogenesis outside complex I deficiency. Third, we will generate induced pluripotent stem cells (iPSCs) from NUBPL patient cells and differentiate neurons from iPSCs to develop a model to study neuronal-specific effects of NUBPL disease and validate efficacies of candidate therapies at neuronal levels.

Utilizing the PS Gene Editing System to Encode a Blood‐Brain Barrier‐Penetrating IDUA as a Treatment for Murine Hurler Syndrome

Awardee: Michael Przybilla

Institution: University of Minnesota

Award Amount: $165,000

Funding Period: February 1, 2023 - January 31, 2024

The new lysosomal activator GHF201 (144DG11) as a potential treatment for mucopolysaccharidosis I”

Awardee: Or Kakhlon

Institution: Research Fund of the Hadassah Medical Organization

Award Amount: $165,000

Funding Period: February 1, 2023 - January 31, 2024

Choroid plexus disruption in CDD

Awardee: Michela Fagiolini

Institution: Boston Children's Hospital

Grant Amount: $50,240.00

Funding Period: February 1, 2023 - January 31, 2024

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Summary:

In the absence of CDKL5 we discovered a significant disruption of Choroid Plexus (ChP) organization at the level of tight junctions and polarity of selective cellular transporters in the epithelial cells. These preliminary results raise the question whether the loss of CDKL5 negatively affects the function of the blood-cerebrospinal fluid (CSF)-barrier and the composition of the CSF. The ChP-CSF system delivers important growth-active molecules throughout the brain, guaranteeing its proper maturation and functioning. Here we propose 1) a detailed characterization of the development of the ChP epithelial cells in the absence of CDKL5 and 2) a multi-omics approach to profile both CSF and ChP in CDKL5 deficient disorder (CDD) mice. Due to the confined but accessible localization of the ChP, our results may establish the ChP as a new tractable target for CDD intervention.

Developing a spatiotemporal gene atlas in iMCD lymph node tissue to identify diagnostic biomarkers and therapeutic targets

Awardee: David Fajgenbaum

Institution: University of Pennsylvania

Grant Amount: $60,570.00

Funding Period: February 1, 2023 - January 31, 2024

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Summary:

Human Herpesvirus (HHV)-8-negative, idiopathic multicentric Castleman disease (iMCD) is a deadly hematologic illness with an unknown etiology that affects individuals of all ages. Although the causes, key immune cell types, signaling pathways, and cytokines involved are poorly understood, a key hallmark of iMCD includes characteristic lymph node features that are used for diagnosis. Despite importance of these lymph node features as diagnostic criteria, the underlying biological mechanisms driving them are not well understood. In the proposed LOI, we will develop a single cell RNAseq/spatial gene expression atlas in iMCD lymph nodes to characterize cell subpopulations, identify cell-type compositional changes, infer intercellular communication networks, and interrogate dysregulated cell types/cell locations within disease tissue. By completing the proposed specific aims we will gain important insights into lymph node biology in iMCD and the thorough characterization of the inflamed tissue will help identify new biomarkers and uncover dysregulated cell types that may be contributing to disease pathogenesis and pathology.

Advance in PIP2 treatment to restore capillary blood flow

Awardee: Fabrice Dabertrand

Institution: University of Colorado Anschutz Medical Campus

Grant Amount: $117,734

Funding Period: February 1, 2023 - January 31, 2024

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Summary:

Neurons in the brain have few energy reserves and therefore depend on local blood flow through arterioles and capillaries for a continuous supply of nutrients. A deficit in cerebral blood flow hemodynamics is an early feature of CADASIL, which suggests that cerebrovascular dysfunction has a key role in the pathogenesis leading to dementia. We recently discovered how to rescue the coupling between neuronal activity and local blood flow regulation using a phospholipid-based treatment with PIP2. However, PIP2 can act on several targets, including as an inhibitor of TRPV4 channels, an important player in capillary blood flow regulation. The proposed work aims at characterizing the impact of CADASIL on this pathway and further develop the PIP2 treatment to improve cerebral blood flow in CADASIL mouse model.

BOOSTING AUTOPHAGY AS A THERAPEUTIC STRATEGY TO TREAT TBC1D24 EPILEPSY

Awardee: Anna Fassio

Institution: University of Genoa

Grant Amount: $103,546.00

Funding Period: February 1, 2023 - January 31, 2024

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Summary:

TBC1D24 has been described by our group and others to play different roles in the brain, however it is not immediate to translate this knowledge in a therapeutic strategy to treat individuals bearing TBC1D24 mutations. We recently identified a degradative cellular process, named autophagy, to be dysfunctional in neuronal cells upon loss of TBC1D24 function. Autophagy is a multistep process responsible for the removal of superfluous or dysfunctional cell components, and it is essential to guarantee neuronal health. Autophagy can be activated by different compounds, and research on small drugs acting on this process is ongoing both in the field of tumors and neurodegenerative diseases. We hypothesize that the use of these new compounds is effective in reverting the autophagy impairment and ameliorates the epileptic phenotype in TBC1D24 patients.

Development of novel gene therapy strategies for treatment of all SCN2A Disorders

Awardee: Samuel Young

Institution: University of Iowa

Grant Amount: $61,068.00

Funding Period: February 1, 2023 - January 31, 2024

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Summary:

SCN2A disorders comprise a complex landscape of both missense and protein-truncating variants, resulting in a diversity of phenotypes that include epilepsy and intellectual disability. Currently, there is no cure for SCN2A Disorders, nor are there methods in development that would provide therapeutic intervention for all forms of SCN2A Disorders. Here, our team proposes proof-of-principle studies that could be beneficial for both missense and protein-truncation cases, providing a single method to treat the entire diversity of SCN2A Disorders.

The central conducting lymphatic system in patients with the Rasopathies Noonan Syndrome and CardioFacioCutaneous Syndrome with and without lymphatic disease

Awardee: Jos M Draaisma

Institution: Radboud University Medical Center

Grant Amount: $60,100.00

Funding Period: February 1, 2023 - January 31, 2024

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Summary:

To get a better insight into the central lymphatic system in adult volunteers with Noonan Syndrome (NS) and CardioFacioCutaneous (CFC) Syndrome without clinical symptoms or signs of lymphatic disease compared to healthy adult volunteers without disease and NS and CFC patients with severe lymphatic disease. This to enable therapy with MEK-inhibitors or lymphovenous anastomosis. The Dutch Noonan Syndrome Foundation participates in this study.

A novel Zebrafish model of Kaposiform Lymphangiomatosis: A tool for drug discovery

Awardee: Karina Yaniv

Institution: Sheba Medical Center

Grant Amount: $68,650.00

Funding Period: February 1, 2023 - January 31, 2024

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Summary:

Kaposiform lymphangiomatosis (KLA) is a member of a broad family of complex lymphatic anomalies (CLA)- rare disorders characterized by the abnormal proliferation of lymphatic vessels in the skin and internal organs. KLA, the most aggressive and rarest form of these disorders, can occur at any age, but the incidence is highest in children and teenagers. Current pharmaceutical treatments are aimed chiefly at managing the symptoms; thus, the 5-year survival rate for children affected by KLA is only about 50%. Therefore, there is an urgent need for new pre-clinical models recapitulating the disease and enabling the identification of novel drug targets. This study aims to characterize a novel KLA zebrafish model we recently established in our lab and to screen for new avenues of treatment. Because of their small size, transparency, and large progeny, ZF have become an attractive means for assessing compound effects at early stages of drug discovery. Recently, a lifesaving treatment for a lymphatic anomaly was identified through a chemical screen based on our early establishment of the ZF as a superb model for the study of lymphatic biology. Here we will harness the power of our novel mutants to screen for compounds that selectively revert the KLA-related phenotypes. We anticipate that completion of this study will help increase our understanding of the etiology of KLA and will lead to the identification of new efficient therapies.

Implementing detection of low-level mosaic variants from blood samples in hyperinsulinism to improve diagnosis

Awardee: Sarah Flanagan

Institution: University of Exeter Medical School

Grant Amount: $70,920.00

Funding Period: February 1, 2023 - January 31, 2024

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Summary:

A genetic diagnosis improves treatment and management of hyperinsulinsm; however, currently ~40% of children referred for genetic testing do not receive a genetic diagnosis. One reason for these missed diagnoses is that some children have mosaic variants, which cannot be detected by standard DNA sequencing. Mosaic variants occur during development so will only be present in some tissues, which is why they are hard to detect in genetic testing of blood samples. Our preliminary data identified 8 patients with previously undetected mosaic variants. Our study will develop these methods for detecting mosaic variants so that they can be included in diagnostic genetic testing for hyperinsulinsm worldwide.

Genetic basis of neuroendocrine cell hyperplasia of infancy

Awardee: Nadia Nathan

Institution: Sorbonne University and Inserm

Grant Amount: $87,145.00

Funding Period: February 1, 2023 - January 31, 2024

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Summary:

Material and methods: Patients will be included in the RespiRare network (Fabre et al. 2022). Patients with a NEHI diagnosis attested by the RespiRare multidisciplinary team (MDT) meeting will be selected. After appropriated consents of the parents, a trio (patient and his two unaffected parents) whole genome sequencing (WGS) will be performed. The identified variants will be studied in terms of in silico pathogenicity and relevance in the context of NEHI pathophysiology. The following gene variants will be selected for comparison between the families: those segregating as new mutations only occurring in affected children and those segregating as recessive traits transmitted from each parent.

Expected results: Identifying molecular causes or predispositions for NEHI is a crucial step in studying the pathophysiology of the disease. This could highlight new pathways of interest that could allow the development of targeted treatments.

Exploring adjuvant immunotherapy to treat lymphangioleiomyomatosis

Awardee: Katharina Maisel

Institution: University of Maryland, College Park

Grant Amount: $75,110.00

Funding Period: February 1, 2023 - January 31, 2024

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Summary:

Lymphangioleiomyomatosis (LAM) is a rare lung diseases that causes cystic destruction of the lungs caused by the abnormal growth of smooth-muscle-like LAM cells that have cancer-like features. To date, rapamycin is the only FDA approved treatment for LAM and this treatment is not a cure. Additionally, about 30% of patients do not respond to the treatment. Therefore, new therapeutic avenues are desperately needed. We and other have recently shown that LAM may cause suppression of the local immune response, similar to cancer, and that re-activating this immune response through checkpoint inhibitor or CAR T cell therapies can enhance survival in a murine model of LAM. Immune adjuvants are another immunotherapy currently under investigation for cancer treatments. We have found that one particular adjuvant, CpG, which activates toll-like receptor 9 (TLR9) on antigen presenting cells can enhance survival in murine LAM. However, this survival is incomplete and thus further investigation is necessary. We have found that repeated dosing of CpG causes an overall reduction of immune cell recruitment to the lungs but does not reduce immunosuppressive regulatory T cells. Repeated TLR stimulation on immune cells can lead to ‘TLR tolerance’, in which the cells become less responsive to the stimulus over time. We hypothesize that TLR tolerance is one of the reasons for incomplete survival after CpG treatment in LAM. Research has also shown that spacing out TLR stimulating treatments or alternating the specific TLR that is stimulated may reduce TLR tolerance. Thus, we will investigate the mechanisms of TLR tolerance in LAM and explore alternative treatments to further increase survival. Overall, this proposal will shed new light onto mechanisms of immunosuppression in LAM and also define new treatment avenues for LAM. Furthermore, this work is the first to use adjuvant immunotherapies as treatments for neoplastic growths with loss of TSC expression and could thus open up the use of these treatments for diseases beyond LAM. Finally, understanding the interplay of immune cells, LAM cells, adjuvant immunotherapies, and loss of TSC expression could lead to new treatment targets/strategies for LAM and other diseases for which adjuvant immunotherapies is used.

Glucan fingerprint as biomarker in Adult polyglucosan body disease (APBD)

Awardee: Berge Minassian

Institution: UT southwestern medical center

Grant Amount: $49,677.00

Funding Period: February 1, 2023 - January 31, 2024

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Summary:

The central paradigm in APBD is excessive and abnormal glycogen in the central nervous system. While it is impossible to non invasively measure this glycogen, we can measure the breakdown products of it in the urine. We devised a technique to profile many of these glycogen breakdown products and showed evidence of being able to use such molecules in the urine of APBD model mice. The current grant will establish testing for other breakdown products in mouse models and patient samples.

Defining APBD pre-clinical biomarkers and assessing a therapy in an APBD mouse model.

Awardee: Matthew Gentry

Institution: University of Florida

Grant Amount: $49,677.00

Funding Period: February 1, 2023 - January 31, 2024

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Summary:

Glycogen is a key energy storage macromolecule in cells. However, the accumulation of aberrant glycogen in the brain drives devastating diseases like Lafora disease (LD) and Adult Polyglucosan Body Disease (APBD). The Gentry laboratory has >15 years of experience defining disease mechanisms for LD and developing pre-clinical therapies and biomarkers that are being translated into the clinic. The Akman laboratory has >15 years of experience defining disease mechanisms for APBD and developing pre-clinical therapies. In this proposal, they will combine efforts to: 1) define the brain metabolic perturbations in an APBD mouse model to identify APBD biomarkers and 2) assess an enzyme therapy as a pre-clinical APBD treatment in the same mouse model. This project brings together two laboratories with non-overlapping expertise in studying glycogen storage diseases to tackle critical questions for the APBD community.

Establishing autophagy inducers as novel therapies in cellular and animal models of Beta-propeller Protein-Associated Neurodegeneration (BPAN)

Awardee: Bertrand Mollereau

Institution: Ecole Normale Supérieure of Lyon

Grant Amount: $69,775.00

Funding Period: February 1, 2023 - January 31, 2024

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Summary:

beta-propeller associated neurodegeneration (BPAN is the most recently identified sub-type of neurodegeneration with brain iron accumulation (NBIA) and there are currently no effective treatments for the disease. BPAN is caused by mutations in an autophagy gene WDR45. Autophagy is an important mechanism regulating neuron survival. Defective autophagy has been observed in several BPAN cellular and models and it was proposed that reduced autophagy could be responsible for neurodegeneration in BPAN patients. Hence, identification of novel therapeutics that restores a functional autophagy constitutes research priority. We have previously identified small molecule compounds that correct autophagy in cultured cells isolated from BPAN patients. We now propose to further develop test and validate the most promising hits. For this purpose, we have developed an animal fly model of BPAN disease exhibiting hallmarks of the disease, such as autophagy defect, iron accumulation, neurodegeneration and locomotor disorder. We will select the best molecule compounds restoring autophagy in human cells to rescue the cellular and locomotor defects them in BPAN flies. From our study, the most promising compounds will then be ready to be tested in a larger animal, with an ultimate aim of clinical translation and tangible patient benefit as soon as possible.

Targeting Metabolic Homeostasis in Dysfunctional CHM Retinal Pigment Epithelia

Awardee: Kathleen Boesze-Battaglia

Institution: University of Pennsylvania

Grant Amount: $61,760.00

Funding Period: February 1, 2023 - January 31, 2024

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Summary:

Loss of vision due to choroideremia (CHM), a progressive retinal degenerative disease affects 1 in 50,000 males. Advanced imaging modalities have recently documented sub-clinical changes in the retinal pigment epithelia (RPE) of CHM patients. While metabolomic studies demonstrate dysfunctional metabolism in CHM patients characterized by a disruption of lipid homeostasis. Collectively, these observations implicate RPE-mediated metabolic dysregulation resulting from loss of Rab Escort Protein-1 (REP1) as an etiological factor in CHM. The Boesze-Battaglia lab has extensive experience with lipid homeostasis in models of human retinal degenerations. To explore the potential of metabolic pathways as therapeutic targets for CHM, we have analyzed induced pluripotent stem cell (iPSC)-derived retinal cell from CHM patients. Treatment strategies for CHM are limited albeit clinical trials for gene augmentation strategies are underway. The efficiency of such treatments may not be truly appreciated or fully assessed for nearly a decade due to the slow progressive nature of the disease. Therefore, there remains an unmet need to explore other options to preserve the health and integrity of the retina prior to noticeable degeneration of the eye. Our goal is to define metabolic imbalance in CHM RPE in an effort to restore metabolic homeostasis and RPE function using CHM-patient specific cell models.

Using TERT variant-to-function analysis to define cryptic telomere biology disorders in sporadic cancers

Awardee: Coleman Lindsley

Institution: Dana-Farber Cancer Institute

Grant Amount: $62,528.00

Funding Period: February 1, 2023 - January 31, 2024

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Summary:

Impaired telomere maintenance is linked to development of cancer. Inherited mutations affecting telomerase cause short telomeres in all tissues and an increased risk of specific cancers. The goal of this proposal is to define the spectrum and functional impact of inherited TERT mutations in adults with cancer and analyze associations with clinical outcomes. To achieve this goal, we have assembled a cohort of 40,000 adult patients with various cancers, will identify TERT variants in existing DNA sequencing data, and perform comprehensive functional interrogation of cancer-associated TERT variants to define their effects on telomerase function.

Lysosomal-Targeted Strategies for TBCK Syndrome

Awardee: Xilma Ortiz-Gonzalez

Institution: University of Pennsylvania

Grant Amount: $40,000.00

Funding Period: February 1, 2023 - January 31, 2024

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Summary:

We have recently co-discovered TBCK-encephaloneuronopathy (TBCKE) syndrome. Many of the patients that helped us establish the genetic link to the disease are my patients in the CHOP neurogenetics clinic. Using patients’ cells, we first reported that TBCK mutations alter autophagy, and then showed that there is secondary mitochondrial dysfunction (ie cellular energy production) in patient cells. We suspect the abnormal mitochondria are due to dysfunctional recycling within the cell, which ultimately happens in the lysosome. Our data suggests that targeting the lysosome (by promoting acidification) can rescue the mitochondrial deficits in TBCK cells. We now propose to further investigate the basis of the lysosomal dysfunction as a potential therapeutic target for TBCK syndrome. We specifically will test 2 strategies that have clinically available (or soon to be available) drugs, to expedite the translation of our preclinical research to future potential clinical trials.

Investigating mechanisms underlying cognitive dysfunction in Glut1 deficiency syndrome

Awardee: Maoxue Tang

Institution: Columbia University

Grant Amount: $61,855.00

Funding Period: February 1, 2023 - January 31, 2024

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Summary:

Glucose Transporter-1 deficiency syndrome (Glut1 DS) is a pediatric-onset brain disorder caused by mutations in one copy (haploinsufficiency) of the SLC2A1 (Glut1) gene and therefore low levels of the SCL2A1-coded protein, Glucose Transporter-1. Patients afflicted with Glut1 DS suffer severe epileptic seizures as children and also exhibit delayed cognitive development. Later in life, a debilitating movement disorder develops and predominates. As of yet, there is no truly effective treatment for Glut1 DS. Moreover, it is unclear how low Glut1 protein causes brain dysfunction. In this project, we wish to understand how low Glut1 results in impaired cognition. We suspect that lactate, a downstream product of brain glucose, is a key mediator of Glut1 DS disease. Low brain glucose in Glut1 DS is thought to reduce levels of brain lactate. Since brain lactate is the preferred energy substrate of cerebral neurons, these neurons are starved. Consequently, they are unable to efficiently connect and communicate with one another. These ideas will be investigated in well-established model mice we have created in the laboratory. At the end of the project, we expect to have a better understanding of how low Glut1 affects cognition and how impairments in cognition correlate with altered brain structure. The project is also expected to identify molecules that rely on adequate brain glucose (and lactate) to ensure that the cerebral circuitry is properly established. Consequently, at the conclusion of this project, we expect to be in a better position to identify therapeutic points of intervention in our quest to treat Glut1 DS effectively and safely.