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The tyrosine kinase inhibitor Nintedanib induces lysosomal dysfunctionality: Role of protonation-dependent

Elena Mosca1, Anja Federa2, Christine Pirker1

  • 1Center of Cancer Research, Comprehensive Cancer Center, Medical University of Vienna, Borschkegasse 8a, 1090, Vienna, Austria.

Chemico-Biological Interactions
|September 16, 2024
PubMed

Insights

Nintedanib (NIN) forms crystalline structures in acidic lysosomes, impacting cell function independently of kinase inhibition. This lysosomal microcrystallization contributes to NIN

Area of Science:

  • Cell Biology
  • Pharmacology
  • Biophysics

Background:

  • Nintedanib (NIN) is a multi-tyrosine kinase inhibitor used for idiopathic pulmonary fibrosis and lung cancer.
  • NIN exhibits protonation-dependent lysosomotropic behavior and unique lysosome-specific fluorescence.
  • The underlying mechanism for these properties and their impact on lysosome function remain unclear.

Purpose of the Study:

  • To investigate the spontaneous formation of Nintedanib matter within acidic cellular compartments.
  • To determine if this matter formation underlies NIN's emissive properties and affects lysosome functionality.
  • To explore the consequences of NIN-induced lysosomal changes on cellular processes and potential adverse effects.

Main Methods:

  • Cellular experiments using Nintedanib and non-protonatable derivatives.
  • Analysis of lysosomal birefringence, fluorescence emission, and vATPase inhibitor sensitivity.
  • In vitro crystallization of NIN under varying pH conditions, coupled with spectroscopic and microscopic analysis.
  • Assessment of lysosomal functionality, autophagy, lipid metabolism, and phospholipidosis in treated cells.
  • In vivo studies in mice orally treated with Nintedanib.

Main Results:

  • Lysosomes treated with NIN, but not its derivatives, showed pH-dependent, spontaneous formation of crystalline NIN structures.
  • These crystalline structures exhibited fluorescence and birefringence, co-localizing within lysosomes.
  • Nintedanib-induced lysosomal crystallization impaired lysosomal function, caused vacuolization, enhanced autophagy, deregulated lipid metabolism, and induced phospholipidosis.
  • Nintedanib also induced phototoxicity dependent on lysosomal microcrystallization.
  • NIN crystalline structures were observed in the gut mucosa of treated mice.

Conclusions:

  • Nintedanib spontaneously forms crystalline matter within acidic lysosomes, leading to unique emissive properties.
  • This kinase inhibition-independent impact on lysosomal functionality drives several of NIN's cell biological activities.
  • Lysosomal microcrystallization of Nintedanib may contribute to its observed adverse effects and phototoxicity.
Keywords:
CrystallizationFluorescenceLysosomotropismNintedanibPhospholipidosisPhototoxicity

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