Filgastrim (granulocyte colony stimulating factor or G-CSF) is a multi-modal hematopoietic growth factor used routinely in the hematology/oncology setting but reports of its benefits for brain diseases are appearing in the literature. Preliminary studies in a mouse model of Alzheimer's disease (AD) in our laboratory suggests it has both pro-cognitive and disease-modifying effects. The mechanisms responsible for these profound effects are not clear and development of G-CSF as a therapeutic agent for patients with AD warrants pre-clinical research. Objectives: The proposed studies are designed to elucidate the cellular and molecular mechanisms by which G-CSF reduces amyloid deposition/levels, stimulates hippocampal neurogenesis and restores synaptic functional activity in a mouse model of AD.
Aim 1 : The hypothesis that enhanced trafficking of bone-marrow derived cells from blood to brain and/or increased microglial and phagocytic activity is responsible for the decrease in beta-amyloid (A-2) deposition will be tested in a chimeric tg AD mouse in which bone marrow derived cells express green fluorescent protein (GFP).
Aim 2 : To test the hypothesis that the pro-cognitive effects of G-CSF are mediated by direct actions on neural progenitor cells and neurons, the extent to which G-CSF promotes neurogenesis and/or promotes recovery of hippocampal synaptic function and structure will be tested with neuroanatomical and electrophysiological techniques.
Aim 3 : At an intracellular level, mechanistic studies will elucidate a novel signaling pathway triggered by G-CSF that involves switching to an alternatively spliced variant of protein kinase C (PKC-delta2) that stimulates hippocampal neurogenesis and inhibits poly- phosphorylation of Tau, both which are postulated to underlie improved hippocampal synaptic function. Methods: Cellular and molecular biology techniques, immunohistochemistry, hippocampal slice electrophysiology and behavioral analysis will be employed. Chimeric tg AD mice engrafted with green fluorescent protein expressing (GFP+) bone marrow cells will be generated to determine the extent to which peripherally derived microglia and other bone marrow-derived cells contribute to amyloid reduction and/or neurogenesis. Findings to date: Administration of G-CSF to cognitively-impaired tg AD mice (APP+PS1) reversed the impaired cognitive performance and significantly decreased A2 deposition in hippocampus and entorhinal cortex. G-CSF-treated tg AD mice also revealed a significant increase in total microgliosis and increased areas of synaptophysin immunostaining in hippocampal CA1 and CA3 regions. Additional preliminary results obtained from G-CSF administration to tg AD mice include stimulation of hippocampal neurogenesis and improvement of electrophysiological function in hippocampal slices. Clinical relevance: G-CSF is routinely and safely used to treat neutropenia and to stimulate hematopoietic stem cell generation in healthy bone marrow donors. The cognitive-enhancing and potential disease-modifying effects of G-CSF in a mouse model of AD provides a strong impetus for clinical trials to reverse or forestall progression of dementia in AD patients. Relevance to VA Mission: AD is an incurable, age-dependent dementing disease with a prevalence of 5 million in the US. As the veteran population ages, the prevalence of AD will increase, resulting in overwhelming demands on the VA health care system. From both humanitarian and socio-economic perspectives, there is an urgent need for disease-modifying medications that delay or reverse the progressive dementia of AD.
Filgastrim (G-CSF) is a multi-modal hematopoietic growth factor, which also has profound effects on the diseased central nervous system. Recent work in our laboratory has demonstrated that administration of G-CSF to cognitively-impaired Alzheimer's disease (AD) transgenic (tg) mice reversed cognitive performance and significantly decreased -amyloid (A ) deposition in hippocampus and entorhinal cortex. G-CSF-treated tg AD mice also revealed a significant increase in total microgliosis and increased areas of synaptophysin immunoreactivity in hippocamal CA1 and CA3 regions. The overall objective of this research project is to elucidate the disease-modifying effects of G-CSF in Tg APP/PS1 mice. We will study events triggered and mediated by G-CSF at the electrophysiological, cellular, and molecular levels that contribute to amyloid reduction, restoration of synaptic activity, hippocampal neurogenesis and cognitive improvement. Given that G-CSF is already utilized clinically to stimulate hematopoietic stem cell production, results from this research project will facilitate translation into clinical trials of G-CSF to reverse or forestall the progression of dementia in AD.
