Motivation
When there is a mismatch between the blood type of an individual and the recipient during blood transfusion, a crucial procedure for medical treatment, serious hemolysis reactions can occur, which can be potentially fatal. In addition to the widely recognized ABO blood type system, the Rhesus D (Rh D) blood type system is also essential and complex, having a significant impact on the safety of blood transfusions. There is a great a significant difference in the percentage of individuals with RhD-negative (RhD-) blood in various parts of the world, with about 14.6% of people in the USA having this type of blood, whereas the percentage is much lower in China, ranging from 0.4% to 1.0%. A variation in this distribution is also seen across a wide range of racial and ethnic groups, highlighting the importance of understanding these differences in order to ensure that blood transfusions are safe and effective. Dedicated to producing universal blood cells (RBCs) for patients of different Rh blood types, this team focuses on generating universal RBCs for diverse patient populations.
Purpose
In this research, we seek to develop universal red blood cells to alleviate blood shortages during emergencies and regular hospital visits. The goal of this initiative is to create red blood cells that can be universally transfused to eliminate the challenges posed by the lack of Rh-negative blood and ensure a steady, readily available supply for all patients. This experiment utilized two distinct approaches, both designed to prevent the detection of Rh antigens by anti-D antibodies. By blocking these antigens, the methods aim to inhibit the activation of the complement system, thereby reducing the risk of potential transfusion failure. These strategies are crucial for improving the safety and efficacy of blood transfusions, particularly in cases involving RhD-negative blood types where compatibility is a critical concern. A practical method is presented for improving the surface of cells for universal blood transfusions, illustrating the potential for rationally designing cell surfaces for transfusion and transplantation. In addition to providing a reliable solution to blood storage and compatibility issues, this advancement could revolutionize emergency care and routine medical procedures.
First chamber (mixing chamber)
Used for thorough mixing of Rh-positive blood and protein. Design an inlet to allow Rh-positive blood to enter, and an outlet channel at the bottom of the chamber to connect to the second chamber. The top of the first chamber has an entrance for inserting a blood bag needle to connect the blood bag and the conversion kit. A blood clot filter is designed at the entrance to filter blood clots and microaggregates, such as broken red blood cells. We chose to place the protein in the first chamber in a solution state to maximize the surface area in contact between the protein and red blood cells.
Second chamber (filtration chamber)
Mix with magnetic beads to filter out red blood cells without attached proteins. Design a built-in filter and place magnetic beads in the filtration chamber. Consider using a circular or rectangular grid to capture unattached red blood cells. Design a one-way valve to ensure that blood can only flow into the third chamber to prevent backflow. Set up a collection outlet, specially designed for red blood cells with attached proteins, to facilitate the flow into the third chamber.
Third chamber (final chamber)
Final processed blood is collected for patient use. The convenient access port is designed to allow medical staff to easily draw blood, and a filter can be installed to prevent impurities from entering. There is a drip chamber at the end for easy observation of blood flow rate.
Future Plan
Year 1: Optimize In Vitro Experiments
Focus: Refine the current in vitro experiments to optimize the anti-D scFv and nanobody binding efficiency.
Plan: Test different concentrations and conditions to improve binding specificity and affinity. Develop assays to measure the effectiveness of Rh D antigen neutralization. Use high-throughput screening to identify the most potent variants. Begin testing for off-target effects and cytotoxicity in human cell lines.
Year 2: Expand In Vitro Studies and Begin In Vivo Feasibility
Focus: Validate in vitro findings and start preliminary in vivo feasibility studies.
Plan: Expand testing to a broader range of cell lines, including those from diverse genetic backgrounds. Develop animal models (e.g., transgenic mice) expressing the human Rh D antigen. Begin initial small-scale in vivo studies to assess safety and potential immune responses. Present findings at a scientific conference and publish in a peer-reviewed journal.
Year 3: In Vivo Optimization and Larger Scale Studies
Focus: Optimize the in vivo approach and expand the scale of animal studies.
Plan: Test multiple dosages, administration routes, and treatment regimens in animal models. Monitor long-term safety, efficacy, and immune response. Use imaging techniques to track the distribution and persistence of anti-D scFv and nanobodies in vivo. Develop a strategy for reducing production costs for wider application.
Year 4: Data Improvement and Refinement of Delivery Mechanisms
Focus: Enhance data quality and refine delivery methods.
Plan: Implement advanced data analysis techniques, like AI-driven modeling, to improve understanding of mechanism and efficacy. Explore alternative delivery methods, such as nanoparticle-based delivery or fusion proteins, for better targeting. Improve experimental designs for higher statistical power and reproducibility. Initiate collaborations with external laboratories to validate results.
Year 5: Introduction to Preclinical Trials and Broaden Public Awareness
Plan: Prepare and submit regulatory documentation for preclinical studies. Engage with regulatory agencies for guidance on clinical development. Begin wider public introduction through webinars, articles in layman-friendly science magazines, and participation in science communication events. Collaborate with stakeholders (e.g., blood banks, hospitals) to assess potential impacts and requirements.
Year 6: Conduct Preclinical Trials and Gather Data for Clinical Application
Focus: Perform preclinical trials and gather extensive data.
Plan: Conduct extensive preclinical trials, focusing on safety, pharmacodynamics, and pharmacokinetics. Work with statisticians to ensure data is robust and interpretable for regulatory submissions. Start preparing for clinical trials by identifying potential partners and funding opportunities. Continue to engage with the public, healthcare professionals, and patient advocacy groups.
Year 7: Start Phase I Clinical Trials and Expand Public Outreach
Focus: Launch Phase I clinical trials and expand awareness.
Plan: Begin Phase I clinical trials to test safety and dosage in a small cohort of healthy volunteers or Rh D-negative individuals. Monitor for any adverse effects and immune responses. Develop educational materials (e.g., infographics, videos) to explain the significance of the research to a broader audience. Publish a comprehensive review of preclinical data in a high-impact journal.
Year 8: Conduct Phase II Clinical Trials and Deepen Public Engagement
Focus: Advance to Phase II trials and deepen public understanding.
Plan: Start Phase II clinical trials to test efficacy in a larger cohort, including individuals with varied genetic backgrounds. Use real-world data to assess broader efficacy and safety profiles. Engage in deeper public communication efforts, including partnerships with media outlets, TED-style talks, and community outreach programs. Apply for grants and funding to support the continuation of clinical trials.
Year 9: Analyze Phase II Results and Plan for Phase III Trials
Focus: Analyze Phase II data and prepare for Phase III.
Plan: Conduct comprehensive data analysis and publish results in scientific and medical journals. Collaborate with international partners to prepare for larger, multicenter Phase III trials. Begin discussions with commercial partners or biotech firms for production and distribution. Organize public forums to discuss trial results and gather feedback from the medical community and the public.
Year 10: Begin Phase III Trials and Prepare for Commercialization
Focus: Start Phase III trials and prepare for market introduction.
Plan: Launch Phase III trials to confirm safety and efficacy across diverse populations. Start working on regulatory submissions for market approval. Collaborate with industry partners for large-scale production and distribution planning. Develop a comprehensive strategy for public education and marketing to introduce the treatment to healthcare providers and patients. This plan offers a structured approach to developing your research into a viable treatment, emphasizing both scientific rigor and public engagement.
