<p>We optimized lipid nanoparticles (LNPs) as a robust delivery system to transport larger therapeutic cargo, such as Prime Editing mRNA, to lung epithelial cells via inhalation. LNPs were chosen over other delivery systems, like Adeno-associated viruses (AAVs), due to their superior cargo capacity and reduced immunogenicity. Our goal was to create a spray-dried lung-specific LNP named AirBuddy, capable of efficiently delivering of our Prime Editing components, referred to as PrimeGuide, to lung tissues through inhalation. This approach is designed to advance precision medicine by ensuring targeted delivery with minimal off-target effects.</p>
<p>We optimized lipid nanoparticles (LNPs) as a robust delivery system to transport larger therapeutic cargo, such as Prime Editing mRNA, to lung epithelial cells via inhalation. LNPs were chosen over other delivery systems, like Adeno-associated viruses (AAVs), due to their superior cargo capacity and reduced immunogenicity. Our goal was to create a spray-dried lung-specific LNP named</p>
<p>capable of efficiently delivering of our Prime Editing components, referred to as PrimeGuide, to lung tissues through inhalation. This approach is designed to advance precision medicine by ensuring targeted delivery with minimal off-target effects.</p>
@@ -221,10 +223,11 @@ export function Description() {
<LoremShort/>
</Collapsible>
<br/>
<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('tabid','/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>, Benjamin Moorlach and the Physical and Biophysical Chemistry working group 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 [1], making our nanoparticles lung-specific. Additionally, we employed a <strong>spray-drying technique</strong> by RNhale [2] to improve the stability of our LNP, ensuring that it withstands the inhalation process without degradation and by that, <strong>AirBuddy</strong> was born. </p>
<p>The SORT LNPs are especially suited for pulmonary delivery due to their capacity for precise organ targeting. Their structural stability is maintained during the delivery process, and the spray-drying approach significantly enhances their resilience, allowing the LNPs to remain intact throughout inhalation. This stability is crucial for the efficient delivery of mRNA into lung epithelial cells, where PrimeGuide can effectively perform genome editing. To evaluate the delivery efficiency, we transfected HEK293 cells using fluorescent cargo and quantified the results through FACS analysis.</p>
<p>To ensure that AirBuddy meets the necessary standards for safety and efficacy, we conducted extensive <aonClick={()=>goToPageAndScroll ('In-Depth Characterization of LNPsH','/materials-methods')}> characterization of the LNPs </a> characterization of the LNPs 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>
<divclassName='row align-items-center'>
<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('tabid','/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>, Benjamin Moorlach and the Physical and Biophysical Chemistry working group 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 [1], making our nanoparticles lung-specific. Additionally, we employed the <strong>spray-drying technique</strong> in cooperation with RNhale [2] to improve the stability of our LNP, ensuring that it 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 HEK293 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>
@@ -302,10 +302,6 @@ export function Engineering() {
<imgsrc="https://static.igem.wiki/teams/5247/engineering-cycle/precysec-casette.svg"alt="PreCyseC modular PE casette"style={{height:"50%",width:"auto"}}/>
</div>
</div>
{/* <H4 text="Test" id="text"/>
<p></p>
<H4 text="Learn" id="text"/>
<p></p> */}
</p>
</div>
<Sectiontitle="References"id="references">
...
...
@@ -574,9 +570,6 @@ export function Engineering() {
<H4text="Conclusion"id="text"/>
We use DMG-PEG2000 in our SORT LNP-based AirBuddy because of its superior biodegradability, enhanced stability, and reduced risk of immune system activation. By building on insights from experts and incorporating principles from Wang’s LNP work, we’ve tailored our nanoparticles for lung-specific delivery. This choice ensures that our formulations remain stable long enough to deliver the therapeutic payload effectively while minimizing potential long-term toxicity. This balance is crucial for pulmonary applications, where DMG-PEG2000 outperforms alternatives like mPEG-2000-DSPE, making it the ideal choice for treating CFTR-related lung diseases.
{/* <H4 text="text" id="text" />
<p>erster absatz</p>
<p>zweiter absatz</p> */}
</p>
</Collapsible>
<p>DMG-PEG2000 of the SORT LNP offers better biodegradability and enhanced stability in pulmonary applications - it is known for its faster clearance and reduced potential for long-term toxicity. To ensure we addressed this issue, cytotoxicity tests were performed in addition to the determination of physical properties in cooperation with the Physical and Biophysical Chemistry working group of Bielefeld University to characterize the LNPs. More details about the composition of the SORT LNPs and function of the components can be read below.</p>
