Modulating MMP Activity in hANOs

The inhibitor for multiple MMP proteases (Actinonin) (Chemical formula C₁₉H₃₅N₃O₅, Enzo, Farmingdale, NY), MMP9 specific inhibitor (Chemical formula C₁₆H₁₇F₂N₃O₃S, Sigma-Aldrich), and MMP13 specific inhibitor (Chemical formula C₂₂H₂₀F₂N₄O₂, Sigma-Aldrich) were used to perform MMP inhibition assay. Actinonin can inhibit the activity of MMP1, MMP2, MMP3, MMP7, MMP8, MMP9, MMP10, MMP12, and MMP13. hANOs were treated with Actinonin (25 μM), MMP9 inhibitor (100 μM), and MMP13 inhibitor (10 μM) twice a day, respectively, from Day 10 (when the organoids started differentiation) to Day 24. MMP activity was measured in the supernatant of hANOs collected on Day 17 and Day 24. The dosages of Actinonin, MMP9 specific inhibitor and MMP13 specific inhibitor were modified based on the manufacturer’s manual and published articles^{57 (link)–59 (link)}.

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Li L., Jiao L., Feng D., Yuan Y., Yang X., Li J., Jiang D., Chen H., Meng Q., Chen R., Fang B., Zou X., Luo Z., Ye X., Hong Y., Liu C, & Li C. (2024). Human apical-out nasal organoids reveal an essential role of matrix metalloproteinases in airway epithelial differentiation. Nature Communications, 15, 143.

Publication 2024

Actinonin

Corresponding Organization :

Other organizations : The First Affiliated Hospital, Sun Yat-sen University, Sun Yat-sen University, Guangzhou Medical University, Guangzhou First People's Hospital, State Key Laboratory of Respiratory Disease, Hainan University

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Variable analysis

independent variables

Actinonin (25 μM)
MMP9 inhibitor (100 μM)
MMP13 inhibitor (10 μM)

dependent variables

MMP activity measured in the supernatant of hANOs collected on Day 17 and Day 24

control variables

HANOs were treated with the inhibitors twice a day, respectively, from Day 10 (when the organoids started differentiation) to Day 24

controls

Positive control: None mentioned
Negative control: None mentioned

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