HMOX1

Researchers have developed a human in vitro model that closely mimics the complexities of radiation-induced lung injury (RILI) and radiation dose sensitivity of the human lung. Using a previously developed microfluidic human Lung Alveolus Chip lined by human lung alveolar epithelial cells interfaced with lung capillary cells to recreate the alveolar-capillary interface in vitro, the researchers recapitulated many of the hallmarks of RILI, including radiation-induced DNA damage in lung tissue, cell-specific changes in gene expression, inflammation, and injury to both the lung epithelial cells and blood vessel-lining endothelial cells. By also evaluating the potential of two drugs to suppress the effects of acute RILI, the researchers demonstrated their model's capabilities as an advanced, human-relevant, preclinical, drug discovery platform. Researchers have developed a human in vitro model that closely mimics the complexities of radiation-induced lung injury (RILI) and radiation dose sensitivity of the human lung. Using a previously developed microfluidic human Lung Alveolus Chip lined by human lung alveolar epithelial cells interfaced with lung capillary cells to recreate the alveolar-capillary interface in vitro, the researchers recapitulated many of the hallmarks of RILI, including radiation-induced DNA damage in lung tissue, cell-specific changes in gene expression, inflammation, and injury to both the lung epithelial cells and blood vessel-lining endothelial cells. By also evaluating the potential of two drugs to suppress the effects of acute RILI, the researchers demonstrated their model's capabilities as an advanced, human-relevant, preclinical, drug discovery platform. The lung is one of the tissues most sensitive to radiation in the human body. People exposed to high radiation doses following nuclear incidents develop radiation-induced lung injury (RILI), which affects the function of many cell types in the lung, causing acute and sustained inflammation, and in the longer term, the thickening and scarring of lung tissue known as fibrosis. RILI also is a common side effect of radiation therapy administered to cancer patients to kill malignant cells in their bodies, and can limit the maximum radiation dose doctors can use to control their tumors, as well as dramatically impair patients' quality of life. Anti-inflammatory drugs given to patients during radiation therapy can dampen the inflammation in the lungs, called pneumonitis, but not all patients respond equally well. This is because RILI is a complex disorder that varies between patients and is influenced by risk factors, such as age, lung cancer state, and other pre-existing lung diseases, and likely the patient's genetic makeup. In the event of nuclear accidents, which usually involve the one-time exposure to much higher doses of radiation, no medical countermeasures are available yet that could prevent and protect against the damage to the lungs and other organs, making this a key priority of the US Food and Drug Administration (FDA). A major obstacle to developing a much deeper understanding of the pathological processes triggered by radiation in the lung and other organs, which is the basis for discovering medical countermeasures, is the lack of experimental model systems that recapitulate how exactly the damage occurs in people. Small animal preclinical models fail to produce key hallmarks of the human pathophysiology and do not mimic the dose sensitivities observed in humans. And although non-human primate models are considered the gold-standard for radiation injury, they are in short supply, costly, and raise serious ethical concerns; they also are not human and sometimes fail to predict responses observed when drugs move into the clinic. Now, a multi-disciplinary research team at the Wyss Institute for Biologically Inspired Engineering at Harvard University and Boston Children's Hospital led by Wyss Founding Director Donald Ingber, M.D., Ph.D., in an FDA-funded project, has developed a human in vitro model that closely mimics the complexities of RILI and radiation dose sensitivity of the human lung. Lung alveoli are the small air sacs where oxygen and CO2 exchange between the lung and blood takes place, and the major site of radiation pneumonitis. Using a previously developed microfluidic human Lung Alveolus Chip lined by human lung alveolar epithelial cells interfaced with lung capillary cells to recreate the alveolar-capillary interface in vitro, the researchers recapitulated many of the hallmarks of RILI, including radiation-induced DNA damage in lung tissue, cell-specific changes in gene expression, inflammation, and injury to both the lung epithelial cells and blood vessel-lining endothelial cells. By also evaluating the potential of two drugs to suppress the effects of acute RILI, the researchers demonstrated their model's capabilities as an advanced, human-relevant, preclinical, drug discovery platform. The findings are published in Nature Communications. "Forming a better understanding of how radiation injury occurs and finding new strategies to treat and prevent it poses a multifaceted challenge that in the face of nuclear threats and the realities of current cancer therapies needs entirely new solutions," said Ingber. "The Lung Chip model that we developed to recapitulatedevelopment of RILI leverages our extensive microfluidic Organ Chip culture expertise and, in combination with new analytical and computational drug and biomarker discovery tools, gives us powerful new inroads into this problem." Ingber is also the Judah Folkman Professor of Vascular Biology at Harvard Medical School and Boston Children's Hospital, and the Hansjörg Wyss Professor of Bioinspired Engineering at the Harvard John A. Paulson School of Engineering and Applied Sciences. Advanced human in vitro model of RILI The human Lung Alveolus Chip is a 2-channel microfluidic culture system in which primary human lung alveolar epithelial cells are cultured in one channel where they are exposed to air as they would be in the lung. They are also interfaced across a porous membrane with primary human lung capillary endothelial cells in the parallel channel that are constantly perfused with a blood-like nutrient medium that contains circulating human immune cells, which also can contribute to radiation responses. This carefully engineered, immunologically active, alveolar-capillary interface also experiences cyclic mechanical movements mimicking actual breathing motions. Importantly, this living breathing Lung Chip can be transiently exposed to clinically relevant doses of radiation, and then investigated for the effects over an extended period of time. When the Lung Alveolus Chip was exposed to increasing doses of radiation, the cell, tissue, and organ-level responses modeled on-chip closely aligned with clinical observations and, importantly, offered new insights into RILI. Following a one-time radiation treatment, the team could observe breaks in the cells' chromosomes with activation of associated DNA repair machinery, whose numbers increased with the amount of radiation applied to the Lung Alveolus Chip. This was paralleled by increased levels of reactive oxygen species that are known to also damage DNA, as well as many other types of molecules. Cells started to increase their size, a phenomenon known as hypertrophy that is commonly seen in alveolar injury in vivo, the barrier comprised by tightly packed endothelial and epithelial cells started to break down and liquid flowed through the endothelial channel accumulated in the epithelial channel. "Interestingly, we mapped the levels of multiple pro-inflammatory cytokines over a seven-day course following the radiation treatment, which is especially important for assessing oncoming radiation injury to the lung. RILI symptoms in patients only begin to appear after a week following exposure, but the preceding inflammation cannot be captured in patients," said first-author Queeny Dasgupta, Ph.D., who led the project as a Postdoctoral Fellow on Ingber's team, and now is a Scientist at Systemic Bio, a 3D Systems company. "Once overt RILI has manifested itself in patients, it is often too late to rescue the affected lung tissue." The team found that the first pro-inflammatory cytokines started to be upregulated already 6 hours following the application of high radiation doses, and that their numbers and levels kept increasing until day seven, the end of their observation period. Importantly, this trend was much stronger pronounced in vascular endothelial cells than lung epithelial cells. In parallel, the cellular injury in the Lung Alveolus Chip was reversed in epithelial cells over time but, in contrast, it was sustained in endothelial cells, replicating clinical observations that found RILI to predominantly impact the vascular endothelium in alveoli. From radiation injury to genes to targets To more systematically understand the cellular changes triggered by radiation in the Lung Chip, and identify potential drug targets, the researchers analyzed the complete gene expression programs of epithelial and endothelial cells over time. This allowed them to not only further define the cell-specific early and later-stage inflammatory responses, but also to generate whole-genome gene expression data that they could feed into a machine learning-based computational algorithm called "Network Model for Causality-Aware Discovery" (NeMoCAD). NeMoCAD previously enabled Ingber's team to predict therapeutic targets and repurpose drugs that reverse disease states. The analysis led them to home in on a gene called HMOX1, which is involved in an anti-oxidant response, and whose expression was upregulated immediately following radiation exposure and remained elevated throughout the seven-day course of the investigation. "We found that further increasing HMOX1 levels with the drug lovastatin in the Lung Alveolus Chip reduced DNA damage and cellular hypertrophy early after the radiation, similarly to the anti-inflammatory drug prednisolone, which we used as a positive control. Later, however, lovastatin worsened the disruption of the endothelial barrier. In fact, by experimentally knocking down HMOX1 expression during later stages, we could partially reverse its later adverse effects," explained Dasgupta. "This showed that HMOX1 function indeed is very relevant to the development of RILI, but also suggests that targeting HMOX1 and perhaps targets related to other potential processes might require a more balanced therapeutic approach." The study is part of a larger campaign at the Wyss Institute aiming to investigate acute radiation damage across several organs and tissues. The group has previously modeled acute radiation injury in Organ Chip models of intestine and bone marrow as well, and each exhibited a different radiation dose sensitivity that matched that of humans. Ingber's team also speculates that radiation damage in one organ might also affect the function of other organs and plans to address this possibility by microfluidically linking different Organ Chips in the future. They also think that the hypersensitivities of patients predisposed by other lung diseases to radiation could be modeled in personalized Lung Alveolus Chips. Other authors on the study are past and present members of Ingber's team, including Amanda Jiang, Amy Wen, Robert Mannix, Yuncheng Man, Sean Hall, and Emilia Javorsky. The work was funded by the FDA (under grant #75F40119C10098), and the Wyss Institute at Harvard University.

RBT-1 achieved the primary biomarker endpoint, as well as several key clinical outcome endpoints; positive data supports the potential for RBT-1 to improve postoperative outcomes in cardiothoracic surgery- RBT-1 as a preconditioning agent has the potential to provide patient benefits in multiple surgical settings where organ injury during surgery increases the risk of post-operative complications- -Planned pivotal Phase 3 study of RBT-1 to be initiated in June 2023- SOUTHLAKE, Texas, May 7, 2023 /PRNewswire/ -- Renibus Therapeutics® ("Renibus"), a clinical-stage biopharmaceutical company focusing on the prevention and treatment of cardio-renal diseases, today presented positive final results from a Phase 2 trial evaluating RBT-1 (stannic protoporfin/iron sucrose) in patients undergoing elective cardiac surgery during an oral presentation at the 103rd Annual American Association for Thoracic Surgery (AATS) meeting at the Los Angeles Convention Center, taking place May 6-9, 2023 in Los Angeles, CA. "We are thrilled with the final results of RBT-1 in this Phase 2 study, demonstrating clinically meaningful reductions across key clinical endpoints that we believe will significantly improve the patient's journey in cardiac surgery," said Frank Stonebanks, Co-CEO of Renibus. "These data demonstrate that RBT-1 has broad potential as a preconditioning agent across several large cardiothoracic surgery settings. We look forward to initiating our pivotal Phase 3 study later this quarter and progressing one step closer to our goal of bringing RBT-1 to patients." Summary of Final Phase 2 Results: RBT-1 was studied in a multicenter, double-blind, placebo controlled, Phase 2 clinical trial in patients undergoing coronary artery bypass graft (CABG) and/or cardiac valve surgery on cardiopulmonary bypass. The trial enrolled 152 patients from cardiac surgery centers in the US, Canada and Australia, 132 patients were evaluable for the primary endpoint (ITT population) and 121 were evaluable for clinical outcomes (mITT population), as pre-specified in the statistical analysis plan. RBT-1 treated patients demonstrated a highly significant increase in anti-inflammatory and antioxidant biomarkers of cytoprotective preconditioning, which was the primary endpoint of the study. The biomarkers assessed were interleukin-10 (IL-10), heme oxygenase-1 (HO-1) and Ferritin. In addition, several clinically meaningful improvements were observed. These results are summarized below: A statistically significant (p<0.0001) increase in biomarkers of cytoprotective preconditioning, including IL-10, HO-1, and Ferritin, which achieved the trial's primary endpoint. A statistically significant (-2.7 days, p=0.0243) reduction in days in the ICU. A clinically meaningful reduction in ventilator time (1 day) and hospital length of stay (1.3 days) A statistically significant reduction in 30-day cardiopulmonary readmission rates (-72%, p=0.0391) and 60- and 90-day cardiopulmonary readmission rates (-75% for both, p=0.0189) A clinically meaningful reduction in atrial fibrillation (-34%), troponin I levels (-49%), and MAKE30 (-36%). In a post hoc analysis of a composite endpoint of death, ICU days, 30-day cardiopulmonary readmission rates, atrial fibrillation rates, and hospital days using the win ratio method, a statistically significant benefit was observed among the treated groups (win ratio 1.63, p<0.02). (This statistical method and composite of key clinical outcomes are pre-specified as the primary endpoint in the planned Phase 3 pivotal trial expected to be initiated in June 2023.) RBT-1 treatment also showed reductions vs placebo in rates of hypervolemia (-64%) and anemia (-49%) as reported by the study investigators. RBT-1 demonstrated a generally well-tolerated safety profile in the Phase 2 trial. Renibus is moving forward into its Phase 3 pivotal trial with the 45 mg SnPP/240 mg FeS dose of RBT-1 (administered as a single infusion); there was no statistically significant difference in efficacy between this dose and the 90 mg SnPP/240 mg FeS dose in the Phase 2 trial; however, the lower dose resulted in less photosensitivity. The post hoc win ratio for the 45 mg SnPP/240 mg FeS dose was 2.02 (p=0.004). "Post-operative complications from cardiac surgery represent a major burden for patients as well as an increased strain on healthcare costs. There are currently no approved pharmacological therapies available for use to improve the post-operative course and quality metrics for these patients," said Kevin Lobdell, MD, System Director-Sanger Heart & Vascular Institute, Advocate Health and Clinical Professor Wake Forest University School of Medicine. "RBT-1 has the potential to make a major impact on patients by reducing post cardiac surgery complications through its unique mechanism." Details of the oral presentation at AATS: Title: "A Phase 2 Randomized, Double-Blind, Placebo-Controlled, Multi-Center Trial of RBT-1 Evaluating Cytoprotective Biomarkers & Post-Operative Outcomes in Patients Undergoing Elective Coronary Artery Bypass Graft and/or Valve Surgery on Cardiopulmonary Bypass" Session: CALS and Rescue Date and Time: May 7, 2023, 7:45 am PDT Presenter: Dr. Andre Lamy, MD, MSc, Cardiac Surgeon, Population Health Research Institute, McMaster University, Canada Following conclusion of ATTS, the slides related to the oral presentation will be available on the publications page of the Renibus website at . About the Completed Phase 2 Trial of RBT-1 The Phase 2 study (NCT04564833), which was completed in February 2023, was a randomized, double-blind, multi-center, placebo-controlled trial evaluating the effect of RBT-1 in patients undergoing elective coronary artery bypass graft (CABG) and/or cardiac valve surgery. Renibus announced final results from this study in May 2023, which support the advancement of RBT-1 into a pivotal Phase 3 study. Renibus previously announced interim results from this RBT-1 Phase 2 study in November 2022 and June 2022, and 30-day topline results in January 2023. About Renibus Renibus is a clinical stage biopharmaceutical company dedicated to treating, improving and extending patients' lives by developing breakthrough products to prevent disease progression, improve outcomes and protect against organ damage associated with cardiorenal diseases and complex surgeries. The Company has developed a robust portfolio of products that activate multiple cytoprotective pathways, including organ protection via preconditioning. Renibus' first-in-class lead program, RBT-1 (stannic protoporfin/iron sucrose), is a potent inducer of anti-inflammatory and antioxidant pathways. RBT-1 has finished its Phase 2 development in cardiac surgery and will soon enter a Phase 3 registration study for its lead indication to reduce the risk of post-operative complications following cardiothoracic surgery. RBT-2 is an antioxidant and anti-fibrotic drug that has been shown to reduce the risk of CKD progression in preclinical models and will be in IND enabling and clinical development in 2023. RBT-3, a novel, low molecular weight iron nanoparticle, is one component of RBT-1 and is targeted at reducing the risk of cisplatin-induced nephrotoxicity and will be taken into clinical development in 2023. RBT-9 (stannic protoporfin) is a potent anti-inflammatory and antioxidant drug with broad spectrum anti-viral properties. It has been investigated in a 42-patient Phase 2, randomized, placebo-controlled trial in high-risk patients with COVID-19. The data from this trial indicated the RBT-9 has the potential to significantly improve clinical outcomes in life threatening viral infections. Additional pre-clinical work is underway to help inform the clinical development strategy. For more information, please visit the Company's website at and engage with us on LinkedIn. Investor and Media Contact: Amy Conrad Juniper Point [email protected] 858-914-1962 Business Development Contact Frank Stonebanks Co-CEO, Renibus [email protected] SOURCE Renibus Therapeutics

-RBT-1 met the primary biomarker endpoint as well as several key clinical endpoints; positive data supports the potential for RBT-1 to improve postoperative outcomes in cardiothoracic surgery- -RBT-1 treatment demonstrated a statistically significant reduction in ICU days and 30-day cardiopulmonary readmission rates; patients treated with RBT-1 had reduced time on ventilator as well as reduced rates of atrial fibrillation, hypervolemia, acute kidney injury, troponin I levels and anemia, indicating a broad, organ protective effect consistent with the mechanism of action of RBT-1- -SCCM has awarded a STAR Research Achievement Award to the presentation as a recognition of excellence in critical care research- -Planned pivotal Phase 3 study of RBT-1 to be initiated in Q1 2023- SOUTHLAKE, Texas, Jan. 19, 2023 /PRNewswire/ -- Renibus Therapeutics® ("Renibus"), a clinical-stage biotech company focusing on the prevention and treatment of cardio-renal diseases, today announced that an oral presentation will be delivered on the positive results of RBT-1 from a Phase 2 Study at the Society for Critical Care Medicine (SCCM) 2023 Critical Care Congress at the Moscone Center in San Francisco, CA from January 21- 24, 2023. Dr. Andre Lamy, MD, MSc, Cardiac Surgeon, Population Health Research Institute, and Professor, McMaster University, Canada will be presenting on behalf of the study team. Dr. Lamy is one of the key clinical trial investigators and an advisor to Renibus. RBT-1 was studied in a multicenter, double-blind, placebo controlled, Phase 2 clinical trial in patients undergoing coronary artery bypass graft (CABG) and/or cardiac valve surgery on cardiopulmonary bypass. The trial enrolled 152 subjects from cardiac surgery centers in US, Canada and Australia, of which 132 were evaluable for the primary endpoint (ITT population) and 121 were evaluable for clinical outcomes (mITT population), as pre-specified in the protocol and statistical analysis plan. RBT-1 treated patients demonstrated a highly significant increase in anti-inflammatory and antioxidant biomarkers of cytoprotective preconditioning, which was the primary endpoint of the study. The biomarkers assessed were interleukin-10, heme oxygenase-1 and ferritin. In addition, several clinically meaningful improvements were observed. These results are summarized below: A statistically significant (p<0.0001) increase in biomarkers of cytoprotective preconditioning, which achieved the trial's primary endpoint. A statistically significant (-2.7 days, p<0.03) reduction in days in the ICU. A statistically significant (-71%, p<0.05) reduction in 30-day hospital readmission rates due to cardiopulmonary causes and a 60% reduction in all-cause readmissions. A clinically meaningful reduction in days on ventilator (-1 day), atrial fibrillation rates (-37%), troponin I levels (-61%), AKI rates (-10%) and length of hospital stay (-1.3 days). In an analysis of a composite endpoint of death, ICU days, ventilator days, atrial fibrillation rates, hospital days and hospital admission rates using the win ratio method, a highly statistically significant benefit was observed among the treated groups (win ratio 1.63, p<0.02). A further win ratio analysis (composite endpoint) looking at ICU days and 30-day hospital re-admission rates showed a highly statistically significant benefit (win ratio 1.84, p<0.01). RBT-1 treatment also showed reductions vs placebo in rates of hypervolemia (-64%) and anemia (-49%) as reported by the study investigators. RBT-1 has demonstrated a generally well-tolerated safety profile. "RBT-1 has shown improvements across several key parameters in the post-cardiac surgery setting, including reduction in time on ventilator and ICU days as well as hospital readmission rates, which are highly relevant clinical outcomes in this patient population," stated Dr. Andre Lamy, MD, Cardiac Surgeon, Professor of Surgery, McMaster University, Hamilton. "The results of the Phase 2 study of RBT-1 serve as a positive step forward in advancing the treatment paradigm for cardiorenal diseases and I believe RBT-1 has the potential to become a first-in-class product." Details of the oral presentation at SCCM: Title: Phase 2 Study Interim Results of RBT-1 Effect on Postoperative Course in Elective CABG/Valve Surgery Session: Star Research Presentations: Cardiovascular Date and Time: Saturday, January 21, 2023, 4:15 PM – 4:30 PM Presenter: Dr. Andre Lamy, MD, MSc, Cardiac Surgeon, Population Health Research Institute, McMaster University, Canada Following conclusion of SCCM, the slides related to the oral presentation will be available on the publications page of the Renibus website at . About the Phase 2 Trial of RBT-1 The Phase 2 study (NCT04564833) is a randomized, multi-center, placebo-controlled trial evaluating the effect of RBT-1 in patients undergoing coronary artery bypass graft (CABG) and/or cardiac valve surgery. Renibus previously announced interim results from the RBT-1 Phase 2 study in November 2022 and June 2022, and 30-day topline results earlier in January 2023. About Renibus Renibus is a clinical stage biopharmaceutical company dedicated to treating, improving and extending patients' lives by developing breakthrough products to prevent disease progression, improve outcomes and protect against organ damage in cardiorenal diseases. The Company has developed a robust portfolio of products that activate multiple cytoprotective pathways, including organ protection via preconditioning. Renibus' first-in-class lead program, RBT-1 (stannic protoporfin/iron sucrose), is a potent inducer of Nrf2, IL-10, and ferritin. RBT-1 is currently in Phase 2 development in cardiac surgery and will soon enter a Phase 3 registration study for its lead indication to reduce the risk of postoperative complications following cardiothoracic surgery. RBT-2 is an antioxidant and anti-fibrotic drug that has been shown to reduce the risk of CKD progression in preclinical models and will be in IND enabling and clinical development in 2023. RBT-3, a novel, low molecular weight iron nanoparticle, is one component of RBT-1 and is targeted at reducing the risk of cisplatin-induced nephrotoxicity and will be taken into clinical development in 2023. RBT-9 (stannic protoporfin) is a potent anti-inflammatory and antioxidant drug with broad spectrum anti-viral properties. It has been investigated in a 42-patient Phase 2, randomized, placebo-controlled trial in high-risk patients with COVID-19. RBT-9 significantly improved clinical status by 1.5 points within 7 days of treatment compared with those who received placebo (p=0.035). Improvement was also observed in subjects who were hospitalized at baseline (1.5 point improvement within 7 days, p=0.016). Additionally, in these hospitalized subjects, hospital LOS was reduced by 68.1% (p=0.035) in those treated with RBT-9 compared with placebo. Additional pre-clinical work is underway to help inform the clinical development strategy. For more information, please visit the Company's website at and engage with us on LinkedIn. Investor and Media Contact: Amy Conrad Juniper Point [email protected] 858-914-1962 Business Development Contact Frank Stonebanks Co-CEO, Renibus [email protected] SOURCE Renibus Therapeutics