OUSD (R&E) CRITICAL TECHNOLOGY AREA(S): Combat Casualty Care OBJECTIVE: This STTR project aims to develop an advanced, high-throughput platform using human-derived organoids to predict compound exposure, toxicity, and efficacy. The platform will integrate cutting-edge organoid technologies with automated systems for novel compounds testing to create a comprehensive, physiologically relevant model for safety and effectiveness assessment. DESCRIPTION: Military personnel frequently face various types of trauma in the line of duty (1), including but not limited to blast injuries, burns, fractures, and traumatic brain injuries (TBI). These injuries often result in complex physiological responses that are challenging to study and treat effectively. Inflammation is associated with these disorders and may underlie the risk for health declines (2). This innovative proposal aims to revolutionize the novel drug development by creating a cutting-edge high-through put screening platform that would combine organoid technology (3), microfluidics, automation, and artificial intelligence. Many start-ups are currently working on domain of organoid technology; however, their focus is on tumor related organoids or patient derived organoids. There is a need to develop methods that swiftly and inexpensively produce thousands of organoids for screening purpose. Here, we seek to address critical challenge by requesting an efficient method that has a potential to accelerate the pace of new drug discovery innovation. Traditional 2D cell cultures and animal models have limitations in replicating the complexity of human tissue responses to trauma and potential therapeutic compounds. There is a critical need for a more efficient and accurate screening method to evaluate potential therapeutic compounds for trauma-related injuries such traumatic brain injuries. This will benefit the drug discovery field in facilitating pre-clinical testing of more personalized treatments and will refine studies on the effects of environmental exposures risk to humans. These organoids can not only used for testing new therapeutic targets but can also allow us to expand our knowledge of the molecular mechanisms of environmental exposures. Such a method should be able to mimic the 3D structure and functionality of human tissues while allowing for high-throughput screening of multiple compounds simultaneously (4). This STTR topic will fulfill multiple portfolio areas of DHA (environmental exposures/ traumatic brain injury) and aligns well with the FDA Modernization Act 2.0 (5) by proposing an innovative approach to drug testing that could potentially reduce or eliminate the need for animal testing in certain stages of new drug development. This model will provide a more physiologically relevant model for predicting human responses to novel drugs (such as anti-inflammatory compounds) compared to traditional 2D cell cultures or animal models. Thus, this will help with accelerating the drug development timeline by enabling rapid, high-throughput screening of drug candidates. Current barriers its widespread use, e.g. in routine safety testing are the cost and complexity of organoid culture. PHASE I: The plan for this topic is to develop and optimize protocols for generating and maintaining a diverse array of organoids derived from human and animal (e.g. mouse) pluripotent or adult stem cells. These protocols should be translated to normal and diseased organoids to accurately model military-relevant trauma responses. This would require optimized organoid culture conditions to ensure long-term viability and functionality. Once the organoid conditions are established, this would require designing and fabricating a microfluidic chip capable of housing multiple organoid types. Thus, allowing for the assessment of novel compound effects on multiple organ systems. These two components need to be integrated into automated systems for precise drug administration, real-time monitoring of organoid responses, and data collection. It is important to demonstrate the integration of biosensors to organoid set-up followed by demonstrating an automated data collection and storage systems. Prepare final report and outline plans for Phase II development. PHASE II: The Phase II should consider combined prototype (biosensor and organoid setup) that will require machine learning algorithms to analyze multi-organ response data and generate predictive models for drug toxicity and efficacy. The goal here is to create a user-friendly interface mimicking military relevant trauma such a TBI for researchers to test drug candidate data. This would require an established model showing trauma outcome such as oxidative stress, inflammation etc. This trauma derived organoid will be programmed as fully functional mini-organs, hence will preserve the genetic and epigenetic makeup of diseased state. The organoids will be screened using established therapeutic compounds. Once the tests are completed, analyze, and interpret data from validation studies and then scaling up the system for higher throughput capabilities. It is recommended to optimize the system for long-term studies (up to 30 days) and implement quality control measures throughout the automated processes. Finally developing a validated organoid-driven prototype capable of testing multiple organ systems. In this phase, plan for regulatory documentation and potential submissions along with comprehensive business plan for potential commercialization. Compounds thoroughly tested using this technology have a potential to proceed to clinical trials, upon FDA approval. PHASE III DUAL USE APPLICATIONS: Phase III work is typically oriented towards technology transition to Acquisition Programs of Record and/or commercialization of STTR research or technology. In Phase III, the performer is expected to seek funding from non-STTR government sources and/or the private sector to develop or transition the prototype into a viable product or service for sale in the military or private sector markets. The Phase III description must include the vision or end-state of the research. It must describe one or more specific Phase III military applications and/or supported S&T or acquisition program as well as the most likely path for transition of the STTR from research to operational capability. Additionally, the Phase III section must include (a) one or more potential commercial applications OR (b) one or more commercial technology that could be potentially inserted into defense systems because of this project. For this topic, both military and non-military medical markets should be considered. Realization of a dual-use technology applicable to both the military and civilian use could be achieved via making commercial partners with cell culture-based companies or organoid based start-ups. Military relevant exposures may include compounds/conditions relevant to military environments (chemical warfare agents, extreme stress). Thus the target applications, including drug discovery, as well as fields like organoid technology, toxicology, personalized medicine, and organ development, must be identified at this stage. Similar to the Government sector, organoid platform in the civilian sector could utilize by screening FDA approved drugs for repurposing, toxicity screening of environmental chemicals etc. REFERENCES: 1. Straud CL, Siev J, Messer S, Zalta AK. Examining military population and trauma type as moderators of treatment outcome for first-line psychotherapies for PTSD: A meta-analysis. J Anxiety Disord. 2019 Oct;67:102133. doi: 10.1016/j.janxdis.2019.102133. Epub 2019 Aug 18. PMID: 31472332; PMCID: PMC6739153. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6739153/ 2. Jessica Gill, Hyunhwa Lee, Taura Barr, Tristin Baxter, Morgan Heinzelmann, Hannah Rak, Vincent Mysliwiec, Lower health related quality of life in U.S. military personnel is associated with service-related disorders and inflammation, Psychiatry Research, Volume 216, Issue 1,2014, Pages 116-122, ISSN 0165-1781, https://doi.org/10.1016/j.psychres.2014.01.046. 3. Lampart Franziska L., Iber Dagmar , Doumpas Nikolaos Organoids in high-throughput and high-content screenings. Frontiers in Chemical Engineering, 5, 2023 https://www.frontiersin.org/journals/chemical-engineering/articles/10.3389/fceng.2023.1120348 4. Zhou JQ, Zeng LH, Li CT, He DH, Zhao HD, Xu YN, Jin ZT, Gao C. Brain organoids are new tool for drug screening of neurological diseases. Neural Regen Res. 2023 Sep;18(9):1884-1889. doi: 10.4103/1673-5374.367983. PMID: 36926704; PMCID: PMC10233755. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10233755/ 5. Zushin PH, Mukherjee S, Wu JC. FDA Modernization Act 2.0: transitioning beyond animal models with human cells, organoids, and AI/ML-based approaches. J Clin Invest. 2023 Nov 1;133(21):e175824. doi: 10.1172/JCI175824. PMID: 37909337; PMCID: PMC10617761. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10617761/ KEYWORDS: Organoids, high-throughput screening, tissue, inflammation, trauma, Biofluidic, algorithms
