Amyotrophic lateral sclerosis (ALS) is a motor neuron disorder characterized by the degeneration and death of motor neurons. Currently there is no cure for ALS. Most ALS cases are sporadic. Only 5 to 10 percent of ALS cases are inherited. About 20 percent of familial ALS were caused by genetic mutations in superoxide dismutase 1 (SOD1). Mice expressing SOD1 mutants have been used frequently as models of familial ALS. In SOD1 mutants of different biochemical characteristics, we have shown endothelial degeneration in the spinal cord, microhemorrhages and blood-spinal cord barrier (BSCB) disruption before motor neuron degeneration. Treatment of SOD1G93A mice with a mutant form of activated protein C (APC) with minimal anticoagulant activity, designated as 5A-APC, slows disease progression and extends the lifespan and the symptomatic phase. We have shown that 5A-APC crosses the BSCB and transcriptionally downregulates SOD1 in motor neurons and microglia in SOD1G93A mice. 5A-APC also downregulates SOD1 in cultured N2a differentiated neurons expressing SOD1G85R or SOD1G37R mutants via protease activated receptors 1 and 3, which inhibits nuclear transport of the SP1 transcription factor. In SOD1G93A mice, 5A-APC eliminated microhemorrhages in the spinal cord and BSCB leakage and reduced microglial activation. Protein C (PC) is made as a single polypeptide precursor which undergoes a series of post-translational modifications (PTMs), including vitamin K dependent 3-carboxylation, to form a mature two-chain zymogen which circulates in plasma and can be converted to APC upon thrombin activation. The extensive PTM makes it very challenging to express high levels of recombinant PC in mammalian cells. In this application, two approaches will be employed to enhance the production of fully functional recombinant human 5A-APC in stable CHO cell lines. One approach is to coexpress vitamin K 2,3-epoxide reductase subunit 1 (VKORC1) to increase the efficiency of vitamin K recycling hence the enhancement of 3-carboxylation of 5A-PC. The other approach is to knock down endoplasmic reticulum (ER) stress/chaperone protein GRP78 to increase the efficiency of 5A-PC secretion. The overall goal of the Phase I study is to enhance production of fully functional 5A-APC mutant in mammalian cell system. Once proven successful, the high-producing cell line will be used to develop a manufacturing process in the Phase II study. Upon completion, the Company will seek internal funding to support 5A-APC GMP manufacturing, toxicity studies, IND-filing, clinical phase 1 and 2 studies in ALS patients. Phase 3 clinical study, NDA filing, and commercialization will be done through licensing and strategic partnering.
Public Health Relevance: Currently, there are no effective therapies to prevent or reverse the course of amyotrophic lateral sclerosis (ALS). Recent findings indicated that the compromised blood vessels in the ALS-affected spinal cord and brain areas may play an important role in the disease progression. This application seeks to develop a potential new therapeutic designed to protect blood vessels as well as neurons in the ALS-affected regions.
Thesaurus Terms: 1a,7a-Dihydro-1a-Methyl-7a-(3,7,11,15-Tetramethyl-2-Hexadecenyl)Naphth(2,3-B) Oxirene-2,7-Dione; 2-Amino-6-Trifluoromethoxybenzothiazole; 5,6,7,8-Tetrahydrofolate[{..}]nadp+ Oxidoreductase; Als; Abscission; Acquired Brain Injury; Active Transport, Cell Nucleus; Adverse Effects; Affect; Alteplase; Amyotrophic Lateral Sclerosis; Anti-Inflammatories; Anti-Inflammatory Agents; Anti-Inflammatory; Antibodies; Anticoagulant Agents; Anticoagulant Drugs; Anticoagulants; Antiinflammatories; Antiinflammatory Agents; Apoplexy; Apoptotic; Area; Autoprothrombin Ii; Biochemical; Biosynthetic Proteins; Bleeding; Blood; Blood Coagulation Factor; Blood Coagulation Factor Ix; Blood Coagulation Factor Vii, Activated; Blood Plasma; Blood Vessels; Blood, Cord; Brain; Brain Injuries; C Cell; C Protein; Cbp Protein (Citrate-Binding); Cho Cells; Cell Line; Cell Lines, Strains; Cellline; Cells; Cerebral Stroke; Cerebrovascular Apoplexy; Cerebrovascular Stroke; Cerebrovascular Accident; Cessation Of Life; Chaperone; Characteristics; Chinese Hamster Ovary Cell; Christmas Factor; Clinical; Clinical Research; Clinical Study; Coagulants; Coagulation Factor Ii Receptor; Coagulation Factor Ix; Coagulation Factor Viia; Coagulation Factors; Complex; Cricetinae; Dhfr; Dna Alteration; Dna Mutation; Data; Death; Dehydrogenases; Diabetic Kidney Disease; Diabetic Nephropathy; Dihydrofolate Dehydrogenase; Dihydrofolate Reductase; Dipeptides; Disease; Disease Progression; Disease Model; Disorder; Disulfide Interchange Enzyme; Disulfide Isomerase; Drugs; Ec 1.5.1.3; Ec 3.4.21.22; Erp59 Pdi; Effectiveness; Embryo; Embryonic; Encephalon; Encephalons; Endoplasmic Reticulum; Enzyme Precursors; Enzymes; Epoxides; Epoxy Compounds; Ergastoplasm; Erp59; Excision; Extirpation; Extravasation; F2r; Fda Approved; Factor Ix; Factor Ix Complex; Factor Ix Fraction; Factor Vii, Activated; Factor Viia; Familial Amyotrophic Lateral Sclerosis; Folic Acid Reductase; Fund Raising; Funding; Grp78; Gsbp; Gehrig's Disease; Gene Alteration; Gene Copy Number; Gene Dosage; Gene Mutation; Generations; Genes; Genetic Alteration; Genetic Change; Genetic Defect; Genetic Mutation; Glycoproteins; Glycosylation Site-Binding Protein; Goals; Golgi; Golgi Apparatus; Golgi Complex; Hamsters; Hemorrhage; Hereditary; Hortega Cell; Human; Human, General; Hydroxylation; Inherited; Kidney; Leader Peptide; Leakage; Length Of Life; Licensing; Link; Longevity; Lou Gehrig Disease; Ms (Multiple Sclerosis); Mammalian Cell; Mammals, Hamsters; Mammals, Mice; Man (Taxonomy); Man, Modern; Medication; Medulla Spinalis; Menaquinone; Metabolic Glycosylation; Mice; Micro Rna; Micrornas; Microglia; Microscopic; Modeling; Molecular Chaperones; Motor Cell; Motor Neuron Disease, Amyotrophic Lateral Sclerosis; Motor Neurons; Multiple Sclerosis; Murine; Mus; Mutation; Nerve Cells; Nerve Unit; Nervous System, Brain; Neural Cell; Neurocyte; Neurons; Nuclear Transport; Nucleocytoplasmic Shuttling; Oxidoreductase; Par-1 Receptor; Par1 Receptor; Pdi; Plat; Ptc; Parafollicular Cell; Patients; Peptide Leader Sequences; Pharmaceutic Preparations; Pharmaceutical Agent; Pharmaceutical Preparations; Pharmaceuticals; Pharmacologic Substance; Pharmacological Substance; Phase; Plasma; Plasma Thromboplastin Component; Play; Post-Transcriptional Gene Silencing; Post-Transcriptional Gene Silencings; Post-Translational Modifications; Post-Translational Protein Processing; Posttranscriptional Gene Silencing; Posttranscriptional Gene Silencings; Posttranslational Modifications; Procedures; Production; Proenzymes; Protease-Activated Receptor 1; Protein C; Protein C (Activated); Protein Disulfide Isomerase; Protein Family; Protein Modification; Protein Modification, Post-Translational; Protein Processing, Post-Translational; Protein Processing, Posttranslational; Protein/Amino Acid Biochemistry, Post-Translational Modification; Proteinase-Activated Receptor 1; Proteins; Quelling; Rna Interference; Rna Silencing; Rna Silencings; Rnai; Receptor, Par-1; Recombinant Proteins; Recombinant Tissue Plasminogen Activator; Recombinants; Recycling; Reductases; Removal; Reticuloendothelial System, Blood; Reticuloendothelial System, Serum, Plasma; Riluzole; Risk; Ristocetin Cofactor; Ristocetin-Willebrand Factor; Rodent Model; Role; S-S Rearrangase; Sod-1 Protein; Sod1 Gene Product; Sp1; Sp1 Gene; Safety; Sclerosis, Disseminated; Screening Procedure; Sepsis; Sequence Alteration; Sequence-Specific Posttranscriptional Gene Silencing; Series; Serine Endopeptidases; Serine Protease; Serine Protein Hydrolases; Serine Proteinases; Serum, Plasma; Signal Peptide, Leader; Site; Sodium Channel Blockers; Spillage; Spinal Cord; Stroke; Structure Of Thyroid Parafollicular Cell; Sulfhydryl-Disulfide Interchange Enzyme; Surgical Removal; System; System, Loinc Axis 4; T-Plasminogen Activator; Ttpa; Technology; Testing; Tetrahydrofolate Dehydrogenase; Therapeutic; Thiol-Disulfide Transhydrogenase; Thrombase; Thrombin; Time; Tissue Activator D-44; Tissue Plasminogen Activator; Tissue-Type Plasminogen Activator; Toxic Effect; Toxicities; Transcription Factor Sp1 Gene; Treatment Side Effects; Trypanothione-Glutathione Thioltransferase; Umbilical Cord Blood; Urinary System, Kidney; Variant; Variation; Vascular Accident, Brain; Vitamin K; Vitamin K 2; Vitamin K Quinone; Vitamin K2; Zymogens; Antihemophilic Factor B; Autoprothrombin Iia; Blood Loss; Blood Thinner; Bloodstream Infection; Brain Attack; Brain Damage; Brain Lesion (From Injury); Carboxylate; Carboxylation; Cerebral Vascular Accident; Citrate Carrier; Citrate Periplasmic Carrier Protein; Citrate Transporter; Citrate-Binding Transport Protein; Clotting Factor; Commercialization; Cultured Cell Line; Design; Designing; Disease/Disorder; Disorder Model; Drug/Agent; Dsba Gene Product; Dsba Protein; Dsbc Gene Product; Dsbd Gene Product; Effective Therapy; Endoplasmic Reticulum Stress; Expression Vector; Fetal Cord Blood; Fibrinogenase; Gene Product; Genome Mutation; Gitter Cell; Glycosylation; Improved; Insular Sclerosis; Knock-Down; Life Span; Lifespan; Manufacturing Process; Mesoglia; Mirna; Microglial Cell; Microgliocyte; Motoneuron; Motor Neuron Degeneration; Mouse Model; Mutant; Neuronal; New Therapeutics; Next Generation Therapeutics; Novel Therapeutics; Nucleocytoplasmic Transport; Overexpression; Perivascular Glial Cell; Phase 1 Study; Phase 2 Study; Phylloquinone Epoxide; Phylloquinone Oxide; Polypeptide; Prevent; Preventing; Public Health Relevance; Renal; Resection; Screening; Screenings; Side Effect; Social Role; Stable Cell Line; Stroke; Superoxide Dismutase 1; T-Pa; Therapy Adverse Effect; Thromboplastinogen B; Thrombopoiesis Inhibitor; Transcription Factor; Treatment Adverse Effect; Tricarboxylate Carrier; Tricarboxylate Transporter; Tricarboxylate-Binding C Protein; Vwf; Vascular; Vitamin K Epoxide; Vitamin K(1) Oxide; Vitamin K1 2,3-Epoxide; Vitamin K1 Epoxide; Vitamin K1 Oxide; Von Willebrand Factor; Von Willebrand Protein; Xpra Gene Product
