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<p>To optimize AirBuddy for pulmonary delivery, we collaborated extensively with several experts, including <aonClick={()=>goToPagesAndOpenTab('weber','/human-practices')}>Prof. Weber, Dr. Große-Onnebrink</a> and <aonClick={()=>goToPagesAndOpenTab('kolonkofirst','/human-practices')}>Dr. Kolonko</a> as medical experts, <aonClick={()=>goToPagesAndOpenTab('kristian','/human-practices')}>Prof. Dr. Müller</a>, <aonClick={()=>goToPagesAndOpenTab('radukic','/human-practices')}>Dr. Radukic</a>, <aonClick={()=>goToPagesAndOpenTab('moorlach','/human-practices')}>Benjamin Moorlach</a> and the <aonClick={()=>goToPagesAndOpenTab('biophysik','/human-practices')}>Physical and Biophysical Chemistry working group</a> as academic experts form Bielefeld University and FH Bielefeld as well as <aonClick={()=>goToPagesAndOpenTab('corden','/human-practices')}>Corden Pharma</a> and <aonClick={()=>goToPagesAndOpenTab('rnhale','/human-practices')}>RNhale</a> as industrial experts. Throughout the <aonClick={()=>goToPagesAndOpenTab('delivery head','/engineering')}>development process</a>, we tested two commercially available kits: the <strong>Cayman Chemical LNP Exploration Kit (LNP-102)</strong> and the <strong>Corden Pharma LNP Starter Kit #2</strong>. While the Cayman kit demonstrated limited transfection efficiency, the Corden Pharma formulation significantly enhanced cellular uptake in lung tissues. Building on this, we integrated the <strong>SORT LNP</strong> method based on Wang's research <SupScrollLinklabel="1"/> , making our nanoparticles lung-specific. Additionally, we are employing the <strong>spray-drying technique</strong> in cooperation with RNhale <SupScrollLinklabel="2"/> to improve the stability of our LNP and the <strong>complexation of RNA with chitosan</strong>, ensuring that the LNP and it's cargo withstands the inhalation process without degradation. This stability is crucial for the efficient delivery of mRNA into lung epithelial cells, where PrimeGuide can effectively perform genome editing.</p>
<p>To optimize AirBuddy for pulmonary delivery, we collaborated extensively with several experts, including <aonClick={()=>goToPagesAndOpenTab('weber','/human-practices')}>Prof. Weber, Dr. Große-Onnebrink</a> and <aonClick={()=>goToPagesAndOpenTab('kolonkofirst','/human-practices')}>Dr. Kolonko</a> as medical experts, <aonClick={()=>goToPagesAndOpenTab('kristian','/human-practices')}>Prof. Dr. Müller</a>, <aonClick={()=>goToPagesAndOpenTab('radukic','/human-practices')}>Dr. Radukic</a>, <aonClick={()=>goToPagesAndOpenTab('moorlach','/human-practices')}>Benjamin Moorlach</a> and the <aonClick={()=>goToPagesAndOpenTab('biophysik','/human-practices')}>Physical and Biophysical Chemistry working group</a> as academic experts form Bielefeld University and FH Bielefeld as well as <aonClick={()=>goToPagesAndOpenTab('corden','/human-practices')}>Corden Pharma</a> and <aonClick={()=>goToPagesAndOpenTab('rnhale','/human-practices')}>RNhale</a> as industrial experts. Throughout the <aonClick={()=>goToPagesAndOpenTab('delivery head','/engineering')}>development process</a>, we tested two commercially available kits: the <strong>Cayman Chemical LNP Exploration Kit (LNP-102)</strong> and the <strong>Corden Pharma LNP Starter Kit #2</strong>. While the Cayman kit demonstrated limited transfection efficiency, the Corden Pharma formulation significantly enhanced cellular uptake in lung tissues. Building on this, we integrated the <strong>SORT LNP</strong> method based on Wang's research <SupScrollLinklabel="1"/> , making our nanoparticles lung-specific. Additionally, we are employing the <strong>spray-drying technique</strong> in cooperation with RNhale <SupScrollLinklabel="2"/> to improve the stability of our LNP and the <strong>complexation of mRNA with chitosan</strong>, ensuring that the LNP and it's cargo withstands the inhalation process without degradation. This stability is crucial for the efficient delivery of mRNA into lung epithelial cells, where PrimeGuide can effectively perform genome editing.</p>
<p>To evaluate the <strong>delivery efficiency</strong>, we transfected HEK292 and CFBE41o- cells using fluorescent cargo and quantified the results through FACS analysis. We also ensured that AirBuddy meets the necessary standards for safety and efficacy since we conducted extensive <aonClick={()=>goToPageAndScroll ('In-Depth Characterization of LNPsH','/materials-methods')}> characterization of the LNPs </a>using techniques such as Zeta potential analysis, Dynamic Light Scattering (DLS), Scanning Electron Microscopy (SEM), and Cryogenic Electron Microscopy (cryo-EM). These methods confirmed the uniformity, stability, and optimal size distribution of the nanoparticles. Furthermore, <strong>cytotoxicity assessments</strong> including MTT and proliferation assays demonstrated that our LNPs are biocompatible and do not impede cell growth or function by the incorporation of <aonClick={()=>goToPagesAndOpenTab('it4','/engineering')}>PEG</a> and other ambivalent components. These findings reinforce AirBuddy's potential as a safe and effective tool for pulmonary delivery, with broad implications for gene therapies targeting lung diseases.</p>
<p>We have successfully demonstrated a <b>proof of concept</b> for our gene therapy approach targeting cystic fibrosis. In initial experiments, HEK cells carrying a 3-base deletion analogous to the <i>F508del</i> mutation were transfected with our prime editing complex. The results met our expectations, confirming the viability of our approach for precise gene correction. Based on these findings, we optimized the prime editing complex, leading to the creation of <i>PrimeGuide</i>, a more compact and efficient editing tool. </p>
<p>Central to our <b>delivery system</b> is <b>AirBuddy</b>, a lung-specific lipid nanoparticle designed to stabilize and protect the prime editing complex during transport to lung ionocytes. <i>AirBuddy</i> ensures that the protein complex is delivered specifically to lung cells, enhancing the efficiency of the gene-editing process. By modifying the lipid nanoparticle with protective features, we achieved increased stability, ensuring effective delivery to the target cells. </p>