Meeting Report: The Dallas Consensus Conference on Liver Transplantation for Alcohol Associated Hepatitis

Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/153137/1/lt25681.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/153137/2/lt25681_am.pd

ON01210.Na (Ex-RAD®) mitigates radiation damage through activation of the AKT pathway.

Development of radio-protective agents that are non-toxic is critical in light of ever increasing threats associated with proliferation of nuclear materials, terrorism and occupational risks associated with medical and space exploration. In this communication, we describe the discovery, characterization and mechanism of action of ON01210.Na, which effectively protects mouse and human bone marrow cells from radiation-induced damage both in vitro and in vivo. Our results show that treatment of normal fibroblasts with ON01210.Na before and after exposure to ionizing radiation provides dose dependent protection against radiation-induced damage. Treatment of mice with ON01210.Na prior to radiation exposure was found to result in a more rapid recovery of their hematopoietic system. The mechanistic studies described here show that ON01210.Na manifests its protective effects through the up-regulation of PI3-Kinase/AKT pathways in cells exposed to radiation. These results suggest that ON 01210.Na is a safe and effective radioprotectant and could be a novel agent for use in radiobiological disasters

Identification of ON01210 (Ex-RAD^®) as a radioprotectant from an (E)-Styryl Benzylsulfone chemical library.

<p>(A) Screening protocol used to identify radioprotective compounds from the chemical library. HFL-1 cells were pretreated with varying concentrations of compounds for 24 hours before irradiation (10 Gy), followed by replating into fresh dishes to form colonies. (B) Representative results of cells treated with compounds that show radioprotective activity. (C) ON01210 pretreatment protected HFL-1 cells in a dose responsive manner.</p

ON01210.Na treatment does not inhibit the colony forming potential of human bone marrow cells.

<p>Human bone marrow cells were treated with increasing concentrations of ON01210.Na for 2 hrs (A) or 24 hrs (B), followed by plating into methylcellulose. Total number of CFUs was determined and the average number (n = 3) was determined and plotted along with SEM. (C) Effect of ionizing radiation on the number of colony forming potential of human bone marrow cells.</p

ON01210.Na protects human bone marrow cells from radiation exposure.

<p>Human bone marrow cells were pre-treated with varying concentrations of ON01210.Na (A) or Amifostine (C) and irradiated at 2, 3, and 4 Gy to identify the optimum dose of drug for radioprotection. Varying doses of radiation were used on human bone marrow cells that were pre-treated with 10 uM ON01210.Na (B) or 250 uM Amifostine (D) to calculate the dose reduction factor (DRF). ON01210.Na protected human bone marrow cells with a calculated DRF value of 1.6.</p

ON01210.Na protects murine pluripotent hematopoietic cells.

<p>(A) Colony forming assay of bone marrow cells derived from mice injected with ON01210.Na (500 mg/kg) at 24 hours and 15 minutes prior to whole body irradiation (5.5 Gy). Note the significant increase in colony numbers of cells isolated on day 18 post-IR from ON01210.Na+IR treated mice as compared with vehicle+IR (Control) (***P = <0.001). (B) White blood cell counts of ON01210.Na treated mice showed an increase recovery rate following exposure to IR. (*P = >0.05;**P = <0.05).</p

Protective properties of compounds.

<p>Protective properties of compounds.</p

ON01210.Na treatment in combination with radiation alters the MAPK signaling pathway.

<p>Human Phospho-MAPK antibody arrays were treated with lysates from HFL-1 cells that were pre-treated with Vehicle (A), 20<b> </b>uM ON01210.Na (B), Vehicle plus 10 Gy radiation (C), 20<b> </b>uM ON01210.Na plus 10 Gy radiation (D). ON01210.Na treatment in combination with radiation increased the phosphorylation status of proteins associated with the AKT pathway.</p

Percent recovery of CFU based on schedule.

<p>Percent recovery of CFU based on schedule.</p

ON01210.Na increases PI3K activity in HFL-1 cells and murine bone marrow cells in response to radiation exposure.

<p>Lysates from HFL-1 cells (A) or mouse bone marrow cells (B) were treated with vehicle, increasing concentrations of ON01210.Na or 10<b> </b>mM LiCl for 2 hours and then irradiated with 10 Gy ionizing radiation. Total protein lysates were immunoprecipitated with an anti-PI3 Kinase polyclonal antibody and IP- kinase assays were performed using L-α-Phosphatidylinositol as a substrate. The resulting phospholipids were resolved on TLC plates.</p