CNM-Au8 Gold Nanocatalysis Protects Neurons Against Degeneration and Death in Multiple in vitro Models of Amyotrophic Lateral Sclerosis Karen S. Ho1, Jean-Philippe Richard2,3, Arens Taga2, Michael Bekier4, Alexandre Henriques5, Noëlle Callizot5, Michael T Hotchkin1, Sami J Barmada4, and Nicholas J Maragakis2’. 1Clene Nanomedicine, Salt Lake City, UT; 2Johns Hopkins University, Baltimore MD; 3currently at Reprocell, USA, Inc., Beltsville, MD; 4University of Michigan, Ann Arbor, MI; 5NeuroSys, Gardanne, France. karen@clene.com Introduction – Nanocatalysis CNM-Au8® Catalytic mechanism of action enhances redox state in favor of energy production, while simultaneously lowering cellular oxidative stress Blood-brain barrier penetrant Suspension of 13 nm diameter, catalytically active, clean-surfaced, faceted gold nanocrystals Orally administered No-adverse effect level (NOAEL) nonclinical toxicology findings Well-tolerated; > 300 patient years of clinical exposure Results from Phase 2 Clinical trials presented at this meeting: Posters 034, 035, and 036. Oral P-presentation on RESCUE-ALS Clinical Trial results: Wed., Mar. 16, 11:10 AM Tennessee Ballroom. Acknowledgments We are very grateful to the individuals with ALS and healthy volunteers who donated fibroblasts, without whom the iPSC studies would not have been possible. The exceptional professional support of our colleagues at Clene has been invaluable. This study was funded by Clene Nanomedicine, Inc. (2) cAMP-Mediated CNM-Au8 Motor Neuron Neuroprotection of Wildtype and SOD1 (G93A) Rodent Motor Neurons CNM-Au8Nanocrystal Mechanistic Effects Improved Energy Production and Utilization Neuroprotection, Remyelination (3) CNM-Au8 Neuroprotection of Human C9ORF72 iPSC-Derived Cortical Neurons Objective To determine whether CNM-Au8, a catalytic suspension of clean-surfaced, faceted gold nanocrystals, promotes neuronal survival and function in multiple independent in vitro models of ALS. Methods/Results CNM-Au8’s ability to promote neuronal survival and function in multiple independent in vitro models of ALS: (1) treatment of primary rat spinal motor neurons improves survival, preserves the neurite networks, and reduces cytoplasmic TDP-43 aggregate accumulation after either glutamate excitotoxic injury or exposure to beta-amyloid (Aβ 1-42) oligomers; (2) treatment of spinal motor neurons from transgenic SOD1G93A rats protects motor neurons from death upon exposure to excitotoxic glutamate in a cAMP-dependent manner, and reduces SOD1 protein accumulation in a manner independent of cAMP; (3) treatment of human induced pluripotent stem cell (iPSC)-derived neurons from C9ORF72 patients prevents their death in response to stress caused by mild neurotrophic factor withdrawal. Finally, we show (4) survival and neurite outgrowth of human iPSC-derived motor neurons in co-culture with toxic, SOD1A4V ALS-patient derived astrocytes are substantially and dose-dependently improved with treatment of CNM-Au8. Conclusion Addressing the deficits of ALS with the energetic catalyst CNM-Au8 appears to be a promising new therapeutic strategy for the treatment and disease-modification of ALS. Results (1) CNM-Au8 Neuroprotection of Rodent Spinal Motor Neurons from Glutamate Excitotoxicity and Amyloid-Beta (1-42) Oligomers (4) CNM-Au8 Neuroprotection of Human iPSC-derived Motor Neurons Co-Cultured with Toxic Patient iPSC-Derived Astrocytes Exhibit 99.5