lewis-x-antigen and Amyotrophic-Lateral-Sclerosis

Amyotrophic lateral sclerosis (ALS) is a fatal incurable neurodegenerative disease characterized by progressive degeneration of motor neurons (MNs), leading to relentless muscle paralysis. In the early stage of the disease, MN loss and consequent muscle denervation are compensated by axonal sprouting and reinnervation by the remaining MNs, but this mechanism is insufficient in the long term. Here, we demonstrate that induced pluripotent stem cell-derived neural stem cells (NSCs), in particular the subpopulation positive for LewisX-CXCR4-β1-integrin, enhance neuronal survival and axonal growth of human ALS-derived MNs co-cultured with toxic ALS astrocytes, acting on both autonomous and non-autonomous ALS disease features. Transplantation of this NSC fraction into transgenic SOD1G93A ALS mice protects MNs in vivo, promoting their ability to maintain neuromuscular junction integrity, inducing novel axonal sprouting and reducing macro- and microgliosis. These effects result in a significant increase in survival and an improved neuromuscular phenotype in transplanted SOD1G93A mice. Our findings suggest that effective protection of MN functional innervation can be achieved by modulation of multiple dysregulated cellular and molecular pathways in both MNs and glial cells. These pathways must be considered in designing therapeutic strategies for ALS patients.

Topics: Allografts; Amyotrophic Lateral Sclerosis; Animals; Cell Line; Disease Models, Animal; Humans; Induced Pluripotent Stem Cells; Integrin beta1; Lewis X Antigen; Mice; Mice, Transgenic; Motor Neurons; Muscle, Skeletal; Neural Stem Cells; Receptors, Interleukin-8A; Stem Cell Transplantation; Superoxide Dismutase

Amyotrophic lateral sclerosis (ALS) is a fatal neurological disease characterized by the degeneration of the motor neurons. We tested whether treatment of superoxide dismutase (SOD1)-G93A transgenic mouse, a model of ALS, with a neural stem cell subpopulation double positive for Lewis X and the chemokine receptor CXCR4 (LeX+CXCR4+) can modify the disease's progression. In vitro, after exposure to morphogenetic stimuli, LeX+CXCR4+ cells generate cholinergic motor neuron-like cells upon differentiation. LeX+CXCR4+ cells deriving from mice expressing Green Fluorescent Protein in all tissues or only in motor neurons, after a period of priming in vitro, were grafted into spinal cord of SOD1-G93A mice. Transplanted transgenic mice exhibited a delayed disease onset and progression, and survived significantly longer than non-treated animals by 23 days. Examination of the spinal cord revealed integration of donor-derived cells that differentiated mostly in neurons and in a lower proportion in motor neuron-like cells. Quantification of motor neurons of the spinal cord suggests a significant neuroprotection by LeX+CXCR4+ cells. Both VEGF- and IGF1-dependent pathways were significantly modulated in transplanted animals compared to controls, suggesting a role of these neurotrophins in MN protection. Our results support the therapeutic potential of neural stem cell fractions through both neurogenesis and growth factors release in motor neuron disorders.

Topics: Amyotrophic Lateral Sclerosis; Animals; Axons; Biomarkers; Cell Count; Cell Differentiation; Clone Cells; Disease Models, Animal; Disease Progression; Enzyme-Linked Immunosorbent Assay; Glial Fibrillary Acidic Protein; Immunohistochemistry; Insulin-Like Growth Factor I; Lewis X Antigen; Mice; Mice, Transgenic; Microscopy, Confocal; Motor Neurons; Multipotent Stem Cells; Nerve Regeneration; Receptors, CXCR4; Reverse Transcriptase Polymerase Chain Reaction; Spinal Cord; Superoxide Dismutase; Vascular Endothelial Growth Factor A

Expression of Le(Y), a difucosylated type 2 chain determinant, has been previously identified as a characteristic of cells undergoing apoptosis. Immunohistochemistry using an antibody for Le(Y), as well as nick-end labeling for the detection of DNA breaks, was done on cervical spinal cord sections from ten patients with amyotrophic lateral sclerosis (ALS) and nine patients who had died from other causes. Le(Y)-positive immunoreactivity was seen in the motor neurons of seven ALS cases, but in none of the other cases. Nick-end labeling was also positive in four ALS cases. Double staining of motor neurons by anti-Le(Y) antibody and nick-end labeling was shown in these cases. Other Le(Y)-positive structures, such as reactive astrocytes and fat-laden microglia/macrophages in the lateral and anterior columns, were negative for nick-end labeling. These results suggest that the mechanism of cell death in the spinal motor neurons of ALS may be apoptosis.

Topics: Adult; Aged; Amyotrophic Lateral Sclerosis; Apoptosis; DNA; DNA Damage; Female; Genetic Techniques; Humans; Lewis X Antigen; Male; Middle Aged; Motor Neurons; Spinal Cord