Genes and Genetic Models in Motor Neuron Disorders

描述（由申请人提供）：家族性肌萎缩性侧索硬化症（FALS），“纯”遗传性痉挛性截瘫（HSP）和原发性侧索硬化症（PLS）是由运动神经元变性引起的遗传异质性疾病。这些疾病要么影响上、下运动神经元（FALS），要么主要影响上运动神经元（HSP和PLS）。我的实验室的长期目标是确定遗传对这些疾病发病机制的贡献。这一目标的成功将导致根据疾病机制制定干预措施，以预防、延缓和治疗这些及相关疾病。我们现在计划在过去五年中完成的工作的基础上，作为计划项目（NS 21442）的一部分。我们完成了所有的特定目标，例如我们在染色体2q33上发现了导致隐性FALS （ALS2）和隐性幼型PLS （JPLS1）的ALSIN基因，研究了导致HSP （SPG5）的SPASTIN突变，并在染色体9q21-q22上发现了显性ALS （x染色体）和ALS/痴呆的新位点。我们将进一步扩大隐性ALS和隐性HSP的基因发现工作，开发FALS， JPLS和HSP的遗传模型，并询问致病基因的产物，以了解它们的蛋白质-蛋白质相互作用，以追踪它们的信号通路。我们计划通过三个主要的具体目标来实现这些目标：(1)鉴定染色体15q15-q21上更常见形式的隐性FALS （ALS5）和染色体8q上常见形式的隐性HSP （SPG5A）的基因。(2)建立针对JPLS和ALS表型的alsin和针对HSP表型的spastin的遗传模型（SPG4）。(3)确定alsin的蛋白相互作用。在之前的研究中，我们缩小了ALS5和SPG5A的基因座。我们将使用高通量测序和生物信息学支持来鉴定这些基因，就像我们成功地鉴定了ALS2基因一样。我们将建立alsin和spastin基因敲除模型，研究这些疾病的病理和发病机制。最后，通过双杂交系统和免疫沉淀对alsin和随后的spastin的相互作用伴侣蛋白进行询问。相互作用将通过双标记共聚焦显微镜和FRET分析确认。

DESCRIPTION (provided by applicant): Familial amyotrophic lateral sclerosis (FALS), 'pure' hereditary spastic paraparesis (HSP) and primary lateral sclerosis (PLS) are genetically heterogeneous disorders caused by motor neuron degeneration. These disorders either affect, both the upper and lower motor neurons (FALS), or primarily the upper motor neuron (HSP and PLS). The long-term goal of my laboratory is to identify the genetic contribution to the pathogenesis of these disorders. Success in this goal will lead to the formulation of interventions based on disease mechanisms to prevent, postpone and treat these and related disorders. We now plan to build on the work we accomplished in the last five years as part of a program project (NS 21442). We fulfilled all the Specific Aims such that we identified the ALSIN gene, that cause both recessive FALS (ALS2) and recessive juvenile PLS (JPLS1), on chromosome 2q33, studied mutations in SPASTIN that cause HSP (SPG5), and identified new loci for dominant ALS (on the X-chromosome) and ALS/dementia on chromosome 9q21-q22. We will further expand our gene discovery effort in recessive ALS and recessive HSP develop genetic models of FALS, JPLS, and HSP, and interrogate the products of causative genes for their protein-protein interactions to trace their signaling pathways. We plan to accomplish these goals by three main Specific Aims (1) Identify the genes for a more common form of recessive FALS (ALS5) on chromosome 15q15-q21 and a common form of recessive HSP (SPG5A) on chromosome 8q. (2) Develop genetic models of alsin for the JPLS and ALS phenotypes and of spastin for the HSP phenotype (SPG4). (3) Identify protein interactions of alsin. In the previous period we narrowed the loci for ALS5 and SPG5A. We will use high throughput sequencing and bioinformatic support to identify these genes, as we successfully did the case of the ALS2 gene. We will make knockout models for alsin and spastin, to study the pathology and pathogenesis of these disorders. Finally, the interacting partner proteins of alsin and subsequently of spastin will be interrogated by the two-hybrid systems and immunoprecipitation. Interaction will be confirmed by dual labeled confocal microscopy and FRET analysis.