The objective of this project is to test the hypothesis that a recently identified putative polytopic intramembrane protease (IMPAS1/IMP1) is a critical regulator of brain development. We postulate that IMPAS1 acts via control of a lipid-dependent evolutionary-conserved signaling system. Mutations in two homologous presenilin genes, PS1 and PS2, are the cause of familial Alzheimer's disease (AD). PSs are required for Notch1 receptor-mediated signaling in early development. We and others have recently identified a novel family of diverged proteins (IMPAS (IMP) or SPP/SPPL) structurally related to presenilins. The function of IMP proteins in vivo is unknown. We have found that Ce-imp-2, homologous to human hIMP1, is a critical regulator of a specific development pathway in C. elegans. We recently obtained preliminary data for the generation of knockout mice for mIMP1 and made primary observations that the major phenotypes of the mIMP1 knockout animals are severe brain development defects: exencephaly (brain overgrowth) and anencephaly (brain depletion). In this project we propose to test the following hypotheses. 1) The function of IMP1 is to control neurulation. 2) Inactivation of hIMP1 will lead to brain abnormalities, including neural tube defects and anencephaly, a common cause of perinatal lethality or severe brain pathology in humans. 3) IMPAS proteins are novel essential components of the specific lipid-dependent signaling pathways controlling development. We anticipate that this pathway is connected to Wnt- downstream signaling, and deficiency in IMP1 impairs this signaling leading to spatio-temporal alterations in processes of programmed cell death during a critical stage of neural tube development. The hypotheses will be tested by pursuing the following specific aims:
Aim 1. To determine the role of IMPAS1 in neurulation and development. We will complete the generation of knockout mIMP1-/- mouse strains and will elucidate phenotypes induced by a loss of function of IMP1 in mice. Detailed comparisons of both brain and non-brain phenotype IMP1-deficient animals and wild type animals will be made to predict the putative pathway controlled by IMP1. We will also determine whether exencephaly and anencephaly are related phenotypes; and what genetic-environmental factors may modify the abnormal neurulation in IMP1 knockout animals leading to anencephaly, a common congenital brain defect in humans.
Aim 2. To elucidate molecular pathways regulated by IMP1 gene in vivo. Using molecular signaling assays, examination of spatial-temporal expression of specific markers and transcriptional effectors we will identify whether molecular alterations in IMP1-deficiency animals are similar to those found in animals with impaired signaling essential for normal neurulation and whether IMP1 interact with Wnt- signaling.
Aim 3. To determine whether regulation of programmed cell death is impaired in IMP1-deficient animals. We will determine whether IMP1- deficiency is associated with defects in the processes of programmed cell death, differentiation and proliferation during brain development. We will determine whether IMP1 is important for regulation of apoptosis via Wnt-dependent or independent pathway.
The study of recently identified multipass transmembrane protein (IMPAS1/IMP1) will provide insight into the mechanisms involved in normal and abnormal brain development and regulation of programmed cell death. The gene for the IMP1 protein is structurally related to Alzheimer's disease presenilins. We found in our preliminary data that this protein may be an essential regulator in the formation of the neural tube fundamental event of embryogenesis. We plan to provide evidence for the essential role of this protein in CNS development and signaling in neurulation, which if disrupted, underlies anencephaly, a most common congenital brain defect in humans. ? ?
