Leonardo Barzaghi

Abstract Breast carcinomas are heterogeneous tissues containing cell populations with distinct molecular programs and mechanical states. How this heterogeneity drives collective invasion remains unclear. Tissue fluidization, the shift from a crowded, solid-like state to a more deformable and mobile one, promotes collective motion of cancer cells.

Abstract Why some tumors respond to immunotherapy (“hot” tumors) while others remain resistant (“cold” tumors) is a central challenge in oncology. Elevated RAB5A-dependent endocytosis drives tissue fluidization during the transition to invasive breast carcinoma, but its immunological consequences are unclear. Here we show that RAB5A-driven fluidization induces a mechano-metabolic stress response that disrupts the AMPK–AKAP1–DRP1 mitochondrial fission pathway, causing mitochondrial elongation.

Quantitative RT-PCR Analysis Quantitative RT-PCR analysis was performed as previously described.[3] Total RNA was extracted using the RNeasy Mini kit (Qiagen) and quantified by NanoDrop to assess both concentration and quality of the samples. 1 microgram of RNA was retrotranscribed using “High-Capacity cDNA Reverse Transcription Kit – Thermofisher.” RT minus was made to check the absence of genomic DNA.