Neuronal ceroid lipofuscinosis (NCL) is a genetically heterogeneous group of disorders and is collectively the most common cause of childhood onset neurodegeneration in the U.S. and worldwide. The most prevalent form of NCL onsets in the juvenile years (JNCL, or Batten disease), and is caused by recessive CLN3 mutation. JNCL children suffer from progressive blindness, seizures, phychosis, and cognitive and motor failure, and the disease is invariably fatal. CLN3 encodes a novel transmembrane protein (CLN3p, or battenin) that localizes to the endosomal-lysosomal pathway and is implicated in the regulation of ion homeostasis and vesicular trafficking, but the primary CLN3p function is yet unknown. This proposal is designed to specifically test the hypothesis that CLN3p functions to regulate vesicle acidification through the endosomal pathway, utilizing model systems that include precise genetic mouse and murine neuronal culture models that were engineered to accurately mimic the mutation most commonly associated with the human disease, and importantly, a novel, cutting-edge JNCL patient-derived, induced pluripotent stem (iPS) cell model that has recently been established by this laboratory. The approach that will be undertaken to test the specific hypothesis is: 1) To refine the location and features of the critical CLN3p function in the endocytic pathway, which is proposed to involve vesicular acidification, we will perform cell biological and biochemical assays in our murine neuronal cell culture model, including genetic and pharmacologic perturbation experiments aimed at specifically examining a putative intersection of CLN3p function and chloride channel regulation;2) To systematically identify the regulatory pathways involved in the deficiency in the endocytic pathway in JNCL, we will conduct RNAi screening and mouse genetic studies using an established high-content endocytosis assay and a mouse modeling paradigm, which have the potential to identify putative JNCL disease modifiers. These studies are also likely to identify previously unrecognized regulators of endosomal-lysosomal trafficking, and therefore could broaden our understanding of the role of endocytosis in more common neurodegenerative conditions;3) To test the hypothesis that an early consequence of CLN3p dysfunction in JNCL patient neurons is dysregulated vesicular acidification in the endocytic pathway and that mutation of other regulators of vesicular acidification can also lead to the human disease, we will perform genetic, cell biological and biochemical assays utilizing a collection of patient samples and our innovative CLN3-iPS cell model.

Public Health Relevance

Neuronal ceroid lipofuscinosis (NCL;AKA Batten disease), although classified as a rare inherited disorder, is the most common cause for neurodegeneration among children worldwide and remains poorly understood and without therapeutic treatment options. Because NCL is considered a rare disease, the pharmaceutical industry is unlikely to play a primary role in finding a cure for NCL. The study of endocytosis in NCL will also have broader impact on other more common neurodegenerative diseases that are marked by endocytic pathway involvement, including Alzheimer disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS073813-03
Application #
8445268
Study Section
Developmental Brain Disorders Study Section (DBD)
Program Officer
Morris, Jill A
Project Start
2011-04-01
Project End
2016-03-31
Budget Start
2013-04-01
Budget End
2014-03-31
Support Year
3
Fiscal Year
2013
Total Cost
$355,702
Indirect Cost
$144,608
Name
Massachusetts General Hospital
Department
Type
DUNS #
073130411
City
Boston
State
MA
Country
United States
Zip Code
02199
(2016) Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition). Autophagy 12:1-222
Chandrachud, Uma; Walker, Mathew W; Simas, Alexandra M et al. (2015) Unbiased Cell-based Screening in a Neuronal Cell Model of Batten Disease Highlights an Interaction between Ca2+ Homeostasis, Autophagy, and CLN3 Protein Function. J Biol Chem 290:14361-80
Cotman, Susan L; Mole, Sara E; Kohan, Romina (2015) Future perspectives: Moving towards NCL treatments. Biochim Biophys Acta 1852:2336-8
Mole, Sara E; Cotman, Susan L (2015) Genetics of the neuronal ceroid lipofuscinoses (Batten disease). Biochim Biophys Acta 1852:2237-41
Huber, Robert J; Myre, Michael A; Cotman, Susan L (2014) Loss of Cln3 function in the social amoeba Dictyostelium discoideum causes pleiotropic effects that are rescued by human CLN3. PLoS One 9:e110544
Grubman, A; James, S A; James, J et al. (2014) X-ray fluorescence imaging reveals subcellular biometal disturbances in a childhood neurodegenerative disorder. Chem Sci 5:2503-2516
Lojewski, Xenia; Staropoli, John F; Biswas-Legrand, Sunita et al. (2014) Human iPSC models of neuronal ceroid lipofuscinosis capture distinct effects of TPP1 and CLN3 mutations on the endocytic pathway. Hum Mol Genet 23:2005-22
Sondhi, Dolan; Scott, Emma C; Chen, Alvin et al. (2014) Partial correction of the CNS lysosomal storage defect in a mouse model of juvenile neuronal ceroid lipofuscinosis by neonatal CNS administration of an adeno-associated virus serotype rh.10 vector expressing the human CLN3 gene. Hum Gene Ther 25:223-39
Cotman, Susan L; Karaa, Amel; Staropoli, John F et al. (2013) Neuronal ceroid lipofuscinosis: impact of recent genetic advances and expansion of the clinicopathologic spectrum. Curr Neurol Neurosci Rep 13:366
Hölter, Sabine M; Stromberg, Mary; Kovalenko, Marina et al. (2013) A broad phenotypic screen identifies novel phenotypes driven by a single mutant allele in Huntington's disease CAG knock-in mice. PLoS One 8:e80923

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