Extracellular matrix (ECM) assembly/disassembly is a critical regulator for airway epithelial development and remodeling. Airway organoid is widely used in respiratory research, yet there is limited study to indicate the roles and mechanisms of ECM organization in epithelial growth and differentiation by using in vitro organoid system. Moreover, most of current Matrigel-based airway organoids are in basal-out orientation where accessing the apical surface is challenging. We present a human apical-out airway organoid using a biochemically defined hybrid hydrogel system. During human nasal epithelial progenitor cells (hNEPCs) differentiation, the gel gradually degrade, leading to the organoid apical surfaces facing outward. The expression and activity of ECM-degrading enzymes, matrix metalloproteinases (MMP7, MMP9, MMP10 and MMP13) increases during organoid differentiation, where inhibition of MMPs significantly suppresses the normal ciliation, resulting in increased goblet cell proportion. Moreover, a decrease of MMPs is found in goblet cell hyperplastic epithelium in inflammatory mucosa. This system reveals essential roles of epithelial-derived MMPs on epithelial cell fate determination, and provides an applicable platform enabling further study for ECM in regulating airway development in health and diseases.

细胞外基质 （ECM） 组装/拆卸是气道上皮发育和重塑的关键调节因子。气道类器官广泛用于呼吸研究，但通过使用体外类器官系统来表明 ECM 组织在上皮生长和分化中的作用和机制的研究有限。此外，目前大多数基于 Matrigel 的气道类器官都处于基底方向，难以进入顶端表面。我们提出了一种使用生化定义的混合水凝胶系统的人类顶端气道类器官。在人鼻上皮祖细胞 （hNEPC） 分化过程中，凝胶逐渐降解，导致类器官顶端表面朝外。ECM 降解酶、基质金属蛋白酶（MMP7、MMP9、MMP10 和 MMP13）的表达和活性在类器官分化过程中增加，其中 MMP 的抑制显着抑制正常纤毛，导致杯状细胞比例增加。此外，在炎性粘膜的杯状细胞增生上皮中发现 MMP 减少。该系统揭示了上皮来源的 MMP 在上皮细胞命运决定中的重要作用，并为 ECM 在健康和疾病中调节气道发育提供了一个适用的平台。