<p>Our project has evolved through a deeply collaborative and human-centered approach, integrating diverse feedback from patients, clinicians, researchers, and industry experts. These insights shaped not only the technical aspects of our gene therapy for cystic fibrosis (CF) but also our commitment to addressing real-world patient needs, ethical considerations, and the disparities in CF treatment worldwide. FromMax Beckmann’s[Link] patient perspective to expert guidance on technical and ethical issues, each stakeholder contributed to refining our solution, ensuring it is both innovative and empathetic. Our focus on gene therapy targeting CF’s complex mutations, integrating physiotherapy, and ensuring global accessibility demonstrates our holistic and inclusive vision for this project. Importantly, the collaboration with researchers in nanoparticle stability and gene therapy, along with the development of bilingual surveys and outreach materials, highlights our efforts to make science more accessible and transparent, bridging gaps in knowledge and care. </p>
<p>Our project has evolved through a deeply collaborative and human-centered approach, integrating diverse feedback from patients, clinicians, researchers, and industry experts. These insights shaped not only the technical aspects of our gene therapy for cystic fibrosis (CF) but also our commitment to addressing real-world patient needs, ethical considerations, and the disparities in CF treatment worldwide. From<aonClick={()=>goToPagesAndOpenTab('maxfirst','')}>Max Beckmann’s</a> patient perspective to expert guidance on technical and ethical issues, each stakeholder contributed to refining our solution, ensuring it is both innovative and empathetic. Our focus on gene therapy targeting CF’s complex mutations, integrating physiotherapy, and ensuring global accessibility demonstrates our holistic and inclusive vision for this project. Importantly, the collaboration with researchers in nanoparticle stability and gene therapy, along with the development of bilingual surveys and outreach materials, highlights our efforts to make science more accessible and transparent, bridging gaps in knowledge and care. </p>
<H5text="Human Practices Integration "/>
<p>From the start, we prioritized engaging with CF patients, making sure that our project aligned with both their needs and scientific expectations. Early input from Max Beckmann, a CF patient and friend of our team, guided key design decisions, such as our focus on lung-targeted gene therapy. His insights also shaped aspects like hygiene protocols for immunocompromised patients and the portrayal of CF in our outreach materials. Max’s ongoing feedback provided invaluable emotional insight, helping us ground the project in the real-world experiences of CF patients.</p>
<H5text="Stakeholder Engagement "/>
<p>We consulted with medical professionals like Prof. Dr. Olariu[Link], who emphasized the importance of early diagnosis and mental health support in CF treatment. These insights led us to integrate mental health considerations into our therapy design and focus on reducing racial and global disparities in CF care. Physiotherapist Katrin Westhoff[Link] highlighted the need for a user-friendly inhalation-based therapy for younger patients, validating our direction towards creating accessible treatments. </p>
<p>We consulted with medical professionals like Olariu Timeline <aonClick={()=>goToPagesAndOpenTab('olariu','')}>Dr. Olariu</a> who emphasized the importance of early diagnosis and mental health support in CF treatment. These insights led us to integrate mental health considerations into our therapy design and focus on reducing racial and global disparities in CF care. Physiotherapist <aonClick={()=>goToPagesAndOpenTab('westhoffinv','')}>Katrin Westhoff</a> highlighted the need for a user-friendly inhalation-based therapy for younger patients, validating our direction towards creating accessible treatments. </p>
<H5text="Ethical, Legal, and Regulatory Considerations "/>
<p>Collaboration with regulatory experts, such as Dr. Eva-Maria Berens[Link], ensured our work adhered to ethical and legal standards, particularly in patient consent and biosafety protocols. Through feedback from legal and bioethics committees, we refined our consent processes and improved our management of personal data, ensuring our project complied with both German and international regulations. </p>
<p>Collaboration with regulatory experts, such as <aonClick={()=>goToPagesAndOpenTab('berens','')}>Dr. Eva-Maria Berens</a>, ensured our work adhered to ethical and legal standards, particularly in patient consent and biosafety protocols. Through feedback from legal and bioethics committees, we refined our consent processes and improved our management of personal data, ensuring our project complied with both German and international regulations. </p>
<H5text="Technical and Scientific Adaptations "/>
<p>We continuously integrated expert technical feedback into the project. Contributions from Dr. Benjamin Winkeljann[Link] and Dr. Katharina Kolonko[Link] helped us improve the stability and scalability of our spray-dried lipid nanoparticles (LNPs), which are essential for RNA delivery. This practical focus on scalable solutions allowed us to address global challenges, such as the need for transportable therapies. Collaborating with yeast cultivation experts like Nils Berelsmann[Link], we also optimized RNA delivery systems[Link] to ensure effective lung penetration. </p>
<p>We continuously integrated expert technical feedback into the project. Contributions from <aonClick={()=>goToPagesAndOpenTab('rnhale','')}>Dr. Benjamin Winkeljann</a> and <aonClick={()=>goToPagesAndOpenTab('kolonkofirst','/human-practices')}>Dr. Katharina Kolonko</a> helped us improve the stability and scalability of our spray-dried lipid nanoparticles (LNPs), which are essential for RNA delivery. This practical focus on scalable solutions allowed us to address global challenges, such as the need for transportable therapies. Collaborating with yeast cultivation experts like <aonClick={()=>goToPagesAndOpenTab('nberelsmann','')}>Nils Berelsmann</a>, we also optimized RNA delivery systems[Link] to ensure effective lung penetration. </p>
<H5text="Global Impact and Inclusivity "/>
<p>Recognizing the disparities in CF care across different regions, particularly in underrepresented areas like Asia, we adjusted our approach to create a more inclusive therapy. With feedback from stakeholders like Joshua[Link] from CF Vest International and Dr. Sriram Vaidyanathan[Link], we ensured our therapy addressed a wider range of CF mutations. This global focus led to bilingual surveys and expanded outreach efforts to raise awareness about CF and gene therapy. </p>
<p>Recognizing the disparities in CF care across different regions, particularly in underrepresented areas like Asia, we adjusted our approach to create a more inclusive therapy. With feedback from stakeholders like <aonClick={()=>goToPagesAndOpenTab('joshua','')}>Joshua Bauder</a> from CF Vest International and <aonClick={()=>goToPagesAndOpenTab('sriram','/human-practices')}>Dr. Sriram Vaidyanathan</a>, we ensured our therapy addressed a wider range of CF mutations. This global focus led to bilingual surveys and expanded outreach efforts to raise awareness about CF and gene therapy. </p>
<H5text="Feedback Loops and Project Evolution "/>
<p>We employed a structured feedback cycle based on Gibbs' Reflection Cycle, ensuring our design continuously evolved with stakeholder input. From initial design to public outreach, every phase of PreCyse was shaped by the feedback we received, allowing us to develop a patient-centered and globally relevant solution. </p>
<p>Nils Berelsmann and Hakan Soytürk, specialists in yeast cultivation, played pivotal roles in advancing our project. Nils provided us with a yeast strain compatible with Dr. Saito's[Link] recommendations, particularly one that doesn’t express proteases, which would degrade our nickase candidates. This strain was essential for maintaining the integrity of our engineered enzymes. </p>
<p>Nils Berelsmann and Hakan Soytürk, specialists in yeast cultivation, played pivotal roles in advancing our project. Nils provided us with a yeast strain compatible with <aonClick={()=>goToPagesAndOpenTab('saito','/human-practices')}>Dr. Saito's</a> recommendations, particularly one that doesn’t express proteases, which would degrade our nickase candidates. This strain was essential for maintaining the integrity of our engineered enzymes. </p>
<p>Hakan equipped us with the methodological expertise needed to work with yeast, guiding us through the complexities of yeast cultivation. With their support, we adapted our engineering design specifically for yeast expression. Nils also supplied us with a target vector optimized for yeast expression, which we successfully used to integrate a CasX nickase candidate. This proved especially valuable as our Spu candidates were difficult to integrate due to the use of overhangs. We are still working on this challenge, staying in close contact with the experts to refine our process. </p>
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@@ -402,13 +405,14 @@ function AnalyseMichaela(){
)
}
functionAnalyseBerens(){
const{goToPagesAndOpenTab}=useNavigation();
return(
<Collapsibletitle="Dr. Eva-Maria Berens – Head of ethics committee at University of Bielefeld"id="berensanalyseC">
<p>In our pursuit of generating primary cultures from human nasal epithelial cells for testing, we sought advice from the Biosafety and Security Committee of iGEM, which led us to consult with Dr. Eva-Maria Berens, our ethics officer. Through in-depth discussions with Dr. Berens, we carefully assessed the risks and legal obligations associated with our project. We learned that, given the minimal invasiveness of the procedure, no formal bioethics application was necessary. However, we recognized the importance of developing a legally sound informed consent form for participants, ensuring that we adhered to the relevant legal frameworks. </p>
<p>In collaboration with various legal institutions, we drafted a comprehensive informed consent form that complies with both national regulations in Germany and the specific policies of Bielefeld University. As the first iGEM team to tackle the complex cultivation of human primary nasal epithelial cells, we were committed to paving the way for future teams. To support this, we created a guideline documenting the proper handling of human biomaterial obtained through these types of sample collection. </p>
<p>Additionally, we worked with Ms. Gabriele Anton[Link], the local biodata officer, to establish protocols for the management and storage of sensitive, personally identifiable data. While no special procedures were required for our specific project, we nonetheless made it a priority to implement anonymized methods to protect participant privacy. </p>
<p>The input and feedback from Dr. Berens and Ms. Anton formed the ethical and legal foundation of our project. With their guidance, we were able to proceed with official approval, allowing us to not only test primary cultures with our technology. Moreover, our contributions to biosafety were significantly advanced as we ensured compliance with all legal and ethical standards. </p>
<p>Having established the necessary ethical and legal groundwork, we turned our focus to the practical aspects of handling and cultivating patient samples. This required specialized expertise, which led us to seek assistance fromStemcell Technologies [Link Michaela]to ensure the successful cultivation and maintenance of the nasal epithelial cells. Their support was essential in enabling us to push forward with our testing and bring our project closer to real-world applications. </p>
<p>Having established the necessary ethical and legal groundwork, we turned our focus to the practical aspects of handling and cultivating patient samples. This required specialized expertise, which led us to seek assistance from<aonClick={()=>goToPagesAndOpenTab('michaela','/human-practices')}>Stemcell Technologies</a> ensure the successful cultivation and maintenance of the nasal epithelial cells. Their support was essential in enabling us to push forward with our testing and bring our project closer to real-world applications. </p>
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@@ -479,7 +483,7 @@ function AnalyseMoor(){
return(
<Collapsibletitle="Benjamin Willem Moorlach – Chitosan Expert "id="moorlachanalyseC">
<p>We gained valuable insights into the unique properties of chitosan, a cationic polymer with significant potential to stabilize RNA in our lipid nanoparticle (LNP) formulations. Chitosan offers robust protection against RNases and exhibits heat stability, making it suitable for processing methods like spray drying. Additionally, its mucoadhesive properties enable optimal choice as LNP component. </p>
<p>A critical insight was the necessity for chitosan to be in an acidic environment (pH 4-6) to maintain its positive charge, which is essential for effective RNA interaction. While it cannot replace PEG due to its hydrophilic nature, chitosan is ideal for forming RNA-chitosan complexes, which can then be encapsulated within LNPs. This approach significantly enhances RNA stability during spray drying, a method we intend to further test in collaboration with RNhale[Link]. </p>
<p>A critical insight was the necessity for chitosan to be in an acidic environment (pH 4-6) to maintain its positive charge, which is essential for effective RNA interaction. While it cannot replace PEG due to its hydrophilic nature, chitosan is ideal for forming RNA-chitosan complexes, which can then be encapsulated within LNPs. This approach significantly enhances RNA stability during spray drying, a method we intend to further test in collaboration with <ahref="https://rnhale.com/">Rnhale</a>. </p>
<p> In terms of implementation, Benjamin educated us on the chemical and structural properties of chitosan, reinforcing our approach to improve stability, particularly against heat, in our LNP formulations. He provided guidance on formulating chitosan-RNA complexes and developed a protocol for integrating them into our LNP formulation without affecting the charge of the nanoparticles. Additionally, he supplied us with chitosan in various sizes, enabling us to test different chitosan complexes for optimal results. </p>