Cell-engineered synthetic chylomicrons for oral drug delivery applications

通过口服途径递送生物药物如治疗性肽和蛋白质是一个重要的研究领域。然而，通过任何给药途径的成功肽递送目前是高度挑战性的。肽的口服递送由于其低生物利用度而受到阻碍，这是由于胃肠道（GI）和肝脏中的酶和pH介导的水解导致的低吸收和高首过提取率。鉴于抗糖尿病肽和胰岛素样肽（例如胰高血糖素样肽-1（GLP-1））的成功产生，由于高患者依从性和市场潜力，对开发口服肽递送方法有很大兴趣。乳糜微粒是通过包装新生三酰甘油、胆固醇、胆固醇酯、至少一种脂溶性维生素和载脂蛋白B48（ApoB 48）在肠细胞中形成的脂蛋白（一组可溶性蛋白质，其与血浆中的脂肪或其它脂质联合收割机结合并转运）。乳糜微粒从肠上皮细胞分泌到肠系膜淋巴中。将疏水性分子和药物并入乳糜微粒中可以利用该途径将药物递送至乳糜微粒。在这个项目中，我们的目标是通过劫持和利用肠细胞中的乳糜微粒细胞机制来创建一种新的亲脂性肽递送范式，乳糜微粒实际上充当感兴趣肽的隐形盾牌。通过将基于液滴的微流体技术与机器人技术相结合，我们将能够首次制造合成乳糜微粒（称为乳糜体），使我们能够系统地改变它们的组成并研究支撑其稳定性的分子相互作用。使用这些研究中产生的分子工程规则，我们将设计可以装载用户定义的肽的从头乳糜微粒。使用有机芯片技术结合超分辨率显微镜（纳米显微镜），我们将研究乳糜微粒和肠道内壁之间的相互作用机制。

The delivery of biological drugs such as therapeutic peptides and proteins via the oral route is an important area of research. However, successful peptide delivery via any administration route is currently highly challenging. The oral delivery of peptides is hampered due to their low bioavailability, which results from low absorption and high rates of first-pass extraction, due to enzymatic and pH mediated hydrolysis in the gastrointestinal (GI) tract and liver. Given the successful generation of antidiabetic peptides and insulin-like peptides (such as glucagon-like peptide-1 (GLP-1), there is great interest in developing oral peptide delivery methodologies due to high patient compliance and market potential. Chylomicrons are lipoproteins (a group of soluble proteins that combine with and transport fat or other lipids in the blood plasma) formed in enterocytes via the packaging of nascent triacylglycerols, cholesterol, cholesterol esters, at least one fat soluble vitamin, and apolipoprotein B48 (ApoB48). Chylomicrons are secreted from the enterocyte into the mesenteric lymph. Incorporation of hydrophobic molecules and drugs into chylomicrons can exploit this pathway for drug delivery to the lymphatics. In this project, we aim to create a new paradigm for lipophilic peptide delivery across the gut by hijacking and exploiting chylomicron cellular machinery in enterocytes with the chylomicrons in effect acting as stealth shields for the peptide of interest. By combining droplet-based microfluidics with robotics we will able to manufacture synthetic chylomicrons for the first time (termed chylosomes) enabling us to systematically vary their composition and study the molecular interactions that underpin their stability. Using the resulting molecular engineering rules that will emerge from these studies we will design de-novo chylomicrons that can onboard user-defined peptides. Using organon-a-chip technologies coupled with super-resolution microscopy (nanoscopy) we will then study the mechanism of interaction between the chylomicrons and the gut lining.