https://www.dynotx.com/
Couldn’t this be our AI partner.
Roza Ogurlu from Duke University is presenting an abstract May 14.
https://annualmeeting.asgct.org/program
Just connecting the dots.
“Abstract Body:
Protein misfolding and aggregation in the ER leads to ER stress and triggers the unfolded protein response (UPR) pathway, which in turn can activate the innate immune system. Upon activation, ‘adaptive UPR’ promotes a series of transcriptional and translational events to re-establish ER homeostasis. However, if ER stress persists, an alternate ‘terminal UPR’ program induces apoptosis. Overexpression of Factor VIII (FVIII), a blood clotting factor, and related derivatives following AAV liver gene delivery for treatment of severe hemophilia A has been linked to ER stress induction in preclinical studies and hepatotoxicity in clinical studies. Similarly, other endogenous or exogenous secreted proteins, such as monoclonal antibodies, can elicit ER stress upon over-expression exceeding critical thresholds, thus decreasing efficacy. Consequently, there is an unmet need to circumvent pro-apoptotic UPR, while maintaining sufficient long-term protein expression in mRNA and gene therapies. One strategy is to take advantage of UPR-linked cell survival promoting mechanisms to preserve an optimal rate of protein production from delivered genes.
Here, we exploit an endogenous feedback loop that involves RNase-mediated rearrangement of X-box binding protein 1 (XBP1) mRNA during adaptive UPR to engineer an ER stress responsive switch. To deploy unconventional XBP1 mRNA splicing in gene delivery, we first incorporated XBP1 mRNA fragments (XBP1F1-5) upstream of reporter genes to validate mRNA splicing and protein expression regulation under chemically induced ER stress. We designed the fusion mRNA in a way that XBP1F splicing introduces stop codons preceding the reporter gene sequence. Then, we replaced reporter genes with therapeutically relevant ER stress causing FVIII and Leronlimab (an anti-CCR5 monoclonal antibody being evaluated for HIV treatment). For both therapeutic proteins, we observed efficient splicing of XBP1F constructs and a significant decrease in ER stress markers in various cell lines when expression was modulated by XBP1F splicing. We also demonstrate a potential application for the XBP1F switches in mRNA therapy by evaluating XBP1F splicing efficiency and ER stress marker expression in cells transfected with in vitro transcribed XBP1F1-Leronlimab mRNA. Notably, key structural domains based on XBP1F stem loops were essential for optimal attenuation of ER stress levels. In order to assess this technology in a preclinical proof-of-concept study, we packaged Leronlimab and XBP1F-Leronlimab into AAV8 and administered vectors to mice intravenously. Including XBP1F in the cassette design reduced interindividual variability of therapeutic mRNA expression and ER stress marker levels in the liver.
To conclude, by leveraging an unconventional splicing mechanism during UPR, we developed an RNA-based gene switch to control expression from delivered nucleic acids (mRNA or DNA) encoding ER stress causing proteins. Further testing and refinement of XBP1F modified therapeutic constructs in animal models with the goal of developing mRNA and gene therapies with decreased immunotoxicity and improved safety profiles is underway.”
Summary:
At the upcoming ASGCT (American Society of Gene and Cell Therapy) meeting on May 14, researchers from Duke University, led by Roza Ogurlu, will present major new technology that directly involves leronlimab.
They developed a novel RNA-based gene switch that controls how much protein (like leronlimab) is produced inside cells.
This switch uses the cell’s own unfolded protein response (UPR) to reduce ER stress and prevent toxicity during gene therapy or mRNA therapy delivery.
Critically, they specifically tested leronlimab using this technology:
• Resulted in lower ER stress.
• Stabilized expression levels.
• Reduced immune activation and toxicity.
• Proved effective in cell lines and animal models (mice).
This breakthrough could dramatically improve leronlimab’s safety profile, durability, and effectiveness — opening the door for safer use in mRNA therapies, AAV gene therapies, cancer treatments, autoimmune conditions, and beyond.
Potentially, with the help of AI-based partners like Dyno Therapeutics (who specialize in designing optimal delivery vectors), leronlimab could become part of a new generation of cell and gene therapies with vastly superior performance and safety.
This development has not yet been widely publicized but it represents a critical step forward for Cytodyn’s future, if properly leveraged.
Couldn’t this be our AI partner.
Roza Ogurlu from Duke University is presenting an abstract May 14.
https://annualmeeting.asgct.org/program
Just connecting the dots.
“Abstract Body:
Protein misfolding and aggregation in the ER leads to ER stress and triggers the unfolded protein response (UPR) pathway, which in turn can activate the innate immune system. Upon activation, ‘adaptive UPR’ promotes a series of transcriptional and translational events to re-establish ER homeostasis. However, if ER stress persists, an alternate ‘terminal UPR’ program induces apoptosis. Overexpression of Factor VIII (FVIII), a blood clotting factor, and related derivatives following AAV liver gene delivery for treatment of severe hemophilia A has been linked to ER stress induction in preclinical studies and hepatotoxicity in clinical studies. Similarly, other endogenous or exogenous secreted proteins, such as monoclonal antibodies, can elicit ER stress upon over-expression exceeding critical thresholds, thus decreasing efficacy. Consequently, there is an unmet need to circumvent pro-apoptotic UPR, while maintaining sufficient long-term protein expression in mRNA and gene therapies. One strategy is to take advantage of UPR-linked cell survival promoting mechanisms to preserve an optimal rate of protein production from delivered genes.
Here, we exploit an endogenous feedback loop that involves RNase-mediated rearrangement of X-box binding protein 1 (XBP1) mRNA during adaptive UPR to engineer an ER stress responsive switch. To deploy unconventional XBP1 mRNA splicing in gene delivery, we first incorporated XBP1 mRNA fragments (XBP1F1-5) upstream of reporter genes to validate mRNA splicing and protein expression regulation under chemically induced ER stress. We designed the fusion mRNA in a way that XBP1F splicing introduces stop codons preceding the reporter gene sequence. Then, we replaced reporter genes with therapeutically relevant ER stress causing FVIII and Leronlimab (an anti-CCR5 monoclonal antibody being evaluated for HIV treatment). For both therapeutic proteins, we observed efficient splicing of XBP1F constructs and a significant decrease in ER stress markers in various cell lines when expression was modulated by XBP1F splicing. We also demonstrate a potential application for the XBP1F switches in mRNA therapy by evaluating XBP1F splicing efficiency and ER stress marker expression in cells transfected with in vitro transcribed XBP1F1-Leronlimab mRNA. Notably, key structural domains based on XBP1F stem loops were essential for optimal attenuation of ER stress levels. In order to assess this technology in a preclinical proof-of-concept study, we packaged Leronlimab and XBP1F-Leronlimab into AAV8 and administered vectors to mice intravenously. Including XBP1F in the cassette design reduced interindividual variability of therapeutic mRNA expression and ER stress marker levels in the liver.
To conclude, by leveraging an unconventional splicing mechanism during UPR, we developed an RNA-based gene switch to control expression from delivered nucleic acids (mRNA or DNA) encoding ER stress causing proteins. Further testing and refinement of XBP1F modified therapeutic constructs in animal models with the goal of developing mRNA and gene therapies with decreased immunotoxicity and improved safety profiles is underway.”
Summary:
At the upcoming ASGCT (American Society of Gene and Cell Therapy) meeting on May 14, researchers from Duke University, led by Roza Ogurlu, will present major new technology that directly involves leronlimab.
They developed a novel RNA-based gene switch that controls how much protein (like leronlimab) is produced inside cells.
This switch uses the cell’s own unfolded protein response (UPR) to reduce ER stress and prevent toxicity during gene therapy or mRNA therapy delivery.
Critically, they specifically tested leronlimab using this technology:
• Resulted in lower ER stress.
• Stabilized expression levels.
• Reduced immune activation and toxicity.
• Proved effective in cell lines and animal models (mice).
This breakthrough could dramatically improve leronlimab’s safety profile, durability, and effectiveness — opening the door for safer use in mRNA therapies, AAV gene therapies, cancer treatments, autoimmune conditions, and beyond.
Potentially, with the help of AI-based partners like Dyno Therapeutics (who specialize in designing optimal delivery vectors), leronlimab could become part of a new generation of cell and gene therapies with vastly superior performance and safety.
This development has not yet been widely publicized but it represents a critical step forward for Cytodyn’s future, if properly leveraged.
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