2008 — 2010 |
Borlongan, Cesario V |
U01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Transplantation of Multipotent Progenitor Cells in Stroke @ University of South Florida
Multipotent progenitor cells (MPCs), developed by Dr. Verfaillie and co-workers at the University of Minnesota, appear to have the properties which make them highly suitable as a stem cell therapy. Athersys, Inc. has the proprietary rights to these cells. We have recently developed a collaborative relationship with Athersys, Inc. which has developed manufacturing techniques to make these cells available for clinical use. Athersys, Inc. will supply these cells to us. Our preliminary data in rodents indicate that these cells are quite promising as a therapy for stroke. In order to develop the utility of these cells in stroke, we propose a translational protocol. The proposed set of experiments will serve as the preclinical basis for proceeding with transplantation of MPCs in ischemic stroke patients. Feasibility, safety, and optimal MFC dose and transplantation timing poststroke for producing efficacy in rodent models of adult ischemic stroke will be examined. The following specific aims will serve as milestones, with "go and no-go" criteria. [unreadable] Specific Aim 1: Determine the "long-term" (i.e., 6 months post-transplantation) efficacy of IV rat MPCs in transient middle cerebral artery occlusion model. [unreadable] Specific Aim 2: Evaluate the long-term efficacy of IV human MPCs produced by Athersys, Inc. according to GMP manufacturing protocols in a GLP xenogeneic transient middle cerebral artery occlusion model. [unreadable] Specific Aim 3: Assess the efficacy of IV human MPCs to achieve long-term behavioral benefits in male and female, and adult and aged rats exposed to transient middle cerebral artery occlusion model. [unreadable] Specific Aim 4: Demonstrate the efficacy of IV human MPCs to achieve long-term behavioral benefits in permanent middle cerebral artery ligation model. [unreadable] Specific Aim 5: Characterize stem cell histocompatibility in allogeneic rat MPCs and xenogeneic human MPCs to further provide safety of IV MPCs in rat ischemic stroke model. [unreadable] Specific Aim 6: Conduct IND FDA preparation meetings and assemble documents for filing IND for stem cell therapy in ischemic stroke. This proposal extends our scientific platform for determining the benefit of stem cell therapy in stroke.
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0.958 |
2011 — 2015 |
Borlongan, Cesario V Garbuzova-Davis, Svitlana Nicolai |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Blood-Brain Barrier Repair in Cell Therapy For Stroke @ University of South Florida
DESCRIPTION (provided by applicant): The present proposal advances the motto you break it, we repair it. Recognizing that blood-brain barrier (BBB) breakdown could negatively influence central nervous system (CNS) regenerative processes after stroke, we propose to structurally and functionally restore the BBB in an acute and sub-acute stroke setting. Our preliminary data demonstrate that intravenous administration of a heterogeneous cell population containing stem or progenitor cells shows benefit in animal models of stroke. More recently, we are able to ascribe the functional recovery in transplanted stroke animals to the presence of endothelial progenitor cells (EPC) in the grafted cell population. Whereas cell-based technologies are largely designed to circumvent the BBB for delivery of cells or drugs from the periphery into the brain, we are taking here a novel approach of repairing the BBB damage in stroke. We are also cognizant that the treatment of ischemic stroke is limited to the serine protease tissue-type plasminogen activator (tPA). However, less than 3 percent of ischemic stroke patients benefit from tPA treatment, due to the drug's narrow 3-hour therapeutic window and its detrimental side effects in particular the drug's exacerbation of stroke-induced BBB damage. That 1) stroke is accompanied by BBB damage, 2) tPA adversely contributes to this BBB damage, and 3) cell therapy can afford BBB repair, form the basis of our overarching hypothesis. We posit that any treatment regimen directed at attenuating stroke deficits should consider the pivotal role of BBB repair in order to maintain CNS homeostasis and enhance neuronal regeneration. A regenerative mechanism involving the repair of the damaged BBB by EPC is critical to the successful outcome of cell therapy in stroke, and should also extend the therapeutic window, as well as improve the functional benefits of tPA treatment in stroke.
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0.958 |
2014 — 2015 |
Borlongan, Cesario V |
R21Activity Code Description: To encourage the development of new research activities in categorical program areas. (Support generally is restricted in level of support and in time.) |
Vascular Repair Extends Therapeutic Window For Ischemic Stroke @ University of South Florida
? DESCRIPTION (provided by applicant): On average, one American has stroke every 40 seconds, and one dies every 4 minutes. Of the different types of stroke, acute ischemic stroke is the most common, and successful treatment of this medical condition remains very challenging. The clot busting drug tissue plasminogen activator (tPA) is the only drug approved for clinical use for acute ischemic stroke. However the drug must be initiated within 4.5 h of stroke onset or risk detrimental side effects including intracerebral hemorrhagic transformation (HT). Therefore, an important clinical problem is to develop methods that will extend the limited therapeutic time window of tPA or reduce complications associated with delayed treatment of tPA. The granulocyte-colony stimulating factor (G-CSF) has been shown to exert neuroprotective effects in animal models of ischemia. It is not yet known if the drug could attenuate detrimental side effects of delayed tPA treatment in ischemic stroke. Furthermore, we have shown in a rat model of traumatic brain injury (TBI) that G-CSF monotherapy reduced neuroinflammation in the gray and white matter areas and also ameliorated TBI-induced impairment in endogenous neurogenesis. These findings taken together with reported neuroprotective effects of G-CSF in animal models of ischemia led us to hypothesize that the treatment of G-CSF will also reduce HT associated with delayed treatment of tPA (Aim 1). Treatment with G-CSF mobilizes cells from the bone marrow to the peripheral blood including CD34+ bone marrow stem cells which contain endothelial progenitor cells (EPCs). Several studies suggested that beneficial effects of G-CSF in stroke (e.g. angiogenesis, vasculogenesis, etc.) are mediated by EPCs. Moreover, in a previous study, we have also shown that transplantation of human cerebral endothelial cells attenuated stroke-induced motor and neurological deficits in rats via enhancement of vasculogenesis. In light of these findings, we hypothesized that G-CSF mobilizes EPCs in the setting of tPA-induced HT in stroke, and EPCs attenuate HT via enhancement of vasculogenesis or angiogenesis, processes that preserve the cerebrovasculature (Aim 2). Delayed tPA-induced HT has been attributed to effects of tPA on the neurovascular unit and also via disruption of the blood brain barrier (BBB). We hypothesized that another mechanism underlying neuroprotective effects of G-CSF is via preservation of the integrity of the BBB through vasculogenic and angiogenic effects of recruited EPCs. The long-term goal of this study is to demonstrate that G-CSF in tandem with tPA will reduce delayed tPA-associated complications and also extend the thrombolytic efficacy of tPA. The overall impact is that at the completion of this study, the findings from this work will lay the foundation for the clinical evaluation of G-CSF in attenuating HT associated with delayed treatment of tPA.
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0.958 |
2015 — 2016 |
Borlongan, Cesario V |
R21Activity Code Description: To encourage the development of new research activities in categorical program areas. (Support generally is restricted in level of support and in time.) |
Probing the Stroke Vasculome With Stem Cells @ University of South Florida
? DESCRIPTION (provided by applicant): Stroke survivors exhibit behavioral deficits and brain damage from the primary injury, which can become aggravated over time due to secondary cell death. Each year in the United States, an estimated 795,000 people suffer from stroke and therapeutic interventions are limited with only one FDA-approved drug for ischemic stroke; namely tissue plasminogen activator or tPA. Due to its narrow therapeutic window of only 4.5 hours of ischemic stroke, tPA can only be administered to approximately 5% of ischemic patients due to hemorrhagic complications. Therefore, a safe and effective therapeutic intervention is urgently needed for ischemic stroke patients. It is well recognized that acute and chronic inflammation are associated with stroke pathology. The up regulation of inflammation-associated genes may be linked to the progression of neuro-inflammation after stroke. Recent novel findings suggest that the cerebral endothelium after ischemic injury is no longer passive and can actively upregulate specific genes called stroke vasculome, including Brahma (BRM), I?B (also called NF?B inhibitor), foxfl, and ITIH-5. In addition, the up regulation of non-inflammatory genes in the stroke vasculome also has been detected, such as apcdd1, ATP2b2, and Axin2. Preclinical studies have demonstrated that transplantation of endothelial progenitor cells (EPCs) is a promising treatment because it offers a wider therapeutic window and exerts anti-inflammatory, among other neuroprotective effects, in stroke animals, but no study has directly assessed the potential of EPCs in modifying the inflammation-associated vasculome after ischemic stroke. We advance the hypothesis that anti-inflammatory effects of transplanted EPCs will attenuate the up regulation of inflammatory genes of the stroke vasculome. Our preliminary data indicate that transplantation of human EPCs produced functional recovery primarily by maintaining the integrity of the brain vasculature, specifically preserving the blood brain barrier, which otherwise was leaky after stroke. That EPCs directly exert functional effects on the vasculature gave us the impetus to examine EPC modulation of the inflammation-associated stroke vasculome. Our goal is to better understand the mechanistic role of the vasculome in determining the therapeutic pathway solicited by transplanted EPCs in regulating inflammation within the stroke vasculature. The anticipated outcome that the vasculome (i.e., down regulation of inflammation-associated stroke genes) will reflect efficacy of transplanted EPCs is highly clinically relevant in view of overwhelming clinical findings implicating that vascular risk factors significantly contribute to stroke pathology. To this end, finding a strategy directed at regulating the inflammation-associated vasculome is likely to preserve the integrity of brain vasculature, thereby improving the stroke outcome. Our specific objective in this project is to assess the functional benefit of transplanted EPCs on regulating the inflammation-associated stroke vasculome and to determine the role of specific inflammatory genes in EPC vasculome modulation.
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0.958 |
2015 — 2019 |
Borlongan, Cesario V Garbuzova-Davis, Svitlana Nicolai |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Re-Establishing Vascular Integrity in Als Via Endothelial Cell Transplant @ University of South Florida
? DESCRIPTION (provided by applicant): Amyotrophic lateral sclerosis (ALS) is a fatal disease characterized by widespread motor neuron degeneration in the brain and spinal cord. Mechanisms of disease onset and progression remain poorly understood, but vascular pathology, including damage of the blood-brain barrier (BBB) and blood-spinal cord barrier (BSCB), has recently been recognized as a key factor identifying ALS as a neurovascular disease. Our original studies showed damage to the BBB/BSCB in a mouse model of ALS, characterized by microvessel endothelial cell degeneration in the brain and spinal cord revealed. This new discovery, leading to the innovative concept of vascular leakage in areas of motor neuron degeneration, may point to additional disease mechanisms. Importantly, barrier impairment was noted in ALS mice prior to motor neuron degeneration, suggesting that barrier damage is central to disease initiation. Recent studies, including ours, have shown some degree of barrier damage in ALS patients. Notably, pervasive microvascular barrier impairment was determined in the gray and white matter of the brain and spinal cord from sporadic ALS patients. In this application, we propose to test a new therapeutic approach that has direct relevance to the treatment of ALS. The purpose of this project is to determine whether endothelial progenitor cells from human bone marrow (BMEPCs) can serve as a novel source of transplant cells to repair BSCB integrity and retard motor neuron degeneration in an experimental model of ALS. The significant scientific advance of this project is the demonstration that BMEPC transplantation will restore BSCB competence in ALS and as a result delay motor neuron degeneration. In an effort to determine the optimal transplant regimen, we will assess the effective cell dose then embark on elucidating the reparative processes involved in the repair of BSCB. Whereas most treatment-based studies have largely focused on pre-symptomatic ALS in testing therapeutic efficacy, this proposal strengthens its translational application to the clinic by examining the benefits of BMEPC transplantation in repairing BSCB in a symptomatic mouse model of ALS, essentially equivalent to ALS patients 2-3 years after diagnosis. Specific Aim 1 will establish effect of vascular repair via BMEPC transplant on reversing behavioral deficits (Aim 1A) and motor neuron cell death (Aim 1B) in symptomatic ALS mice. Specific Aim 2 will determine structural vascular repair via BMEPC transplant by examining BMEPC engraftment into the vascular wall (Aim 2A) and microvasculature status of the cervical and lumbar spinal cord with an electron microscope in areas of motor neuron degeneration (Aim 2B). Specific Aim 3 will determine functional vascular repair via BMEPC transplant by assessing endothelial cell viability (Aim 3A), capillary leakage in areas of motor neuron degeneration (Aim 3B), and tightness between endothelial cells (Aim 3C). Our experimental design advances a mechanism-based hypothesis and a highly translational framework for development of a novel therapy for ALS. Positive outcomes from this project will not only provide evidence of a mechanistic role of vascular damage in ALS, but will also offer a preclinical basis to pursue cell therapy to repair the altered barrier. This study represents a relatively low-risk, but high-reward and innovative therapy, as the transplant cells can potentially be expanded from the patient's own bone marrow, thereby expediting entry of this autologous transplantation approach into clinical application for ALS patients.
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0.958 |