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Deciphering the role of CDK4 in regulating metabolism and cell fate of triple-negative breast cancer cells

ABSTRACT: The shift on energetic demands of proliferating cells during tumorigenesis requires an intense crosstalk between cell cycle and metabolism. Beyond their role in cell proliferation, cell cycle regulators also modulate intracellular metabolism of normal tissues. Using both genetic and pharmacological approaches, we aimed to determine the metabolic role of CDK4 in TNBC cells. Unexpectedly, deletion of CDK4 only slightly reduces cell proliferation of triple negative breast cancer (TNBC) cell line and allows in vivo tumor formation. Furthermore, pro-apoptotic stimuli fail to induce proper cell death in CDK4-silenced, depleted or long-term CDK4/6 inhibitors-treated TNBC cells, with dampened mitochondrial calcium uptake. In the third mass spectrometry-based experiment linked to this project, we assayed the proteome and phosphoproteome of purified mitochondria-ER junctions in WT and Cdk4 KO triple negative breast cancer cells (MDA-MB-231).

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Breast Cancer Cell Line, Mammary Gland

DISEASE(S): Breast Cancer

SUBMITTER: Manfredo Quadroni

LAB HEAD: Lluis Fajas

PROVIDER: PXD046327 | Pride | 2024-11-26

REPOSITORIES: pride

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			Action	DRS
	230308_Ziegler_TMT-IMAC_15604.raw	Raw
	230308_Ziegler_TMT-IMAC_15605.raw	Raw
	230308_Ziegler_TMT_15604_2ul.raw	Raw
	230308_Ziegler_TMT_15605_6ul.raw	Raw
	230309_Ziegler_TMT-IMAC_15604_CV40-50.raw	Raw

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Deciphering the role of CDK4 in regulating metabolism and cell fate of triple-negative breast cancer cells

Project description:The shift on energetic demands of proliferating cells during tumorigenesis requires an intense crosstalk between cell cycle and metabolism. Beyond their role in cell proliferation, cell cycle regulators also modulate intracellular metabolism of normal tissues. Using both genetic and pharmacological approaches, we aimed to determine the metabolic role of CDK4 in TNBC cells. Unexpectedly, deletion of CDK4 only slightly reduces cell proliferation of triple negative breast cancer (TNBC) cell line and allows in vivo tumor formation. Furthermore, pro-apoptotic stimuli fail to induce proper cell death in CDK4-silenced, depleted or long-term CDK4/6 inhibitors-treated TNBC cells, with dampened mitochondrial calcium uptake. In the second mass spectrometry-based experiment linked to this project, we used TMT technology to assay the proteome and phosphoproteome of WT and Cdk4 KO cells triple negative breast cancer cells (MDA-MB-231) subjected to mitophagic stress, i.e. a treatment with Oligomycin + Antimycin A.

2024-11-26 | PXD046353 | Pride

Deciphering the role of CDK4 in regulating metabolism and cell fate of triple-negative breast cancer cells

Project description:The shift on energetic demands of proliferating cells during tumorigenesis requires an intense crosstalk between cell cycle and metabolism. Beyond their role in cell proliferation, cell cycle regulators also modulate intracellular metabolism of normal tissues. Using both genetic and pharmacological approaches, we aimed to determine the metabolic role of CDK4 in TNBC cells. Unexpectedly, deletion of CDK4 only slightly reduces cell proliferation of triple negative breast cancer (TNBC) cell line and allows in vivo tumor formation. Furthermore, pro-apoptotic stimuli fail to induce proper cell death in CDK4-silenced, depleted or long-term CDK4/6 inhibitors-treated TNBC cells, with dampened mitochondrial calcium uptake. In the first mass spectrometry-based experiment linked to this project, we explored the proteome and phosphoproteome of WT vs CDK4 KO in triple negative breast cancer cells (MDA-MB-231).

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2024-12-06 | PXD056619 | Pride

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HEK293T cell nuclear & cytosolic proteome remodelling upon amino acid withdrawal. Related to Supplementary Table 4 from An et al (2020)

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CDK4 Inactivation Hinders Cell Death and Metabolism Remodeling Mitochondria-ER contacts in Triple-Negative Breast cancer cells

Project description:The shift in the energetic demands of proliferating cells during tumorigenesis requires extensive crosstalk between the cell cycle and metabolism. Beyond their role in cell proliferation, cell cycle regulators also modulate intracellular metabolism in normal tissues. However, in the context of cancer, where CDK4 is upregulated or stabilized, the metabolic role of CDK4 is barely understood. Here, using both genetic and pharmacological approaches, we sought to determine the metabolic role of CDK4 in triple-negative breast cancer (TNBC) cells. Unexpectedly, the deletion of CDK4 only modestly reduced TNBC cell proliferation and did not hinder tumor formation in vivo. Furthermore, proapoptotic stimuli failed to induce appropriate cell death in TNBC cells upon CDK4 depletion or long-term CDK4/6 inhibitor treatment. Mechanistically, CDK4 enhances mitochondria-endoplasmic reticulum contact (MERC) formation, thereby promoting mitochondrial fission and ER-mitochondrial calcium signaling. Phosphoproteomic analysis also revealed a role for CDK4 in regulating PKA activity at MERCs to sustain ER-mitochondrial calcium signaling. Such CDK4-mediated mitochondrial calcium signaling is required for the metabolic flexibility of TNBC cells. Taken together, these results demonstrate that CDK4 inhibition leads to cell death resistance by inhibiting mitochondrial apoptosis and functions through attenuated MERCs formation and ER-mitochondrial calcium signaling in TNBC. Overall, this study provides new insights into the mechanisms of TNBC resistance to CDK4/6i therapy and paves the way to explore potential synergistic therapeutic targeting of MERCs-associated metabolic shifts.

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