Acid Sphingomyelinase Downregulation Enhances Mitochondrial Fusion and Promotes Oxidative Metabolism in a Mouse Model of Melanoma

Coazzoli, Marco; Napoli, Alessandra; Roux-Biejat, Paulina; Palma, Clara De; Moscheni, Claudia; Catalani, Elisabetta; Zecchini, Silvia; Conte, Vincenzo; Giovarelli, Matteo; Caccia, Sonia; Procacci, Patrizia; Cervia, Davide; Clementi, Emilio; Perrotta, Cristiana

doi:10.3390/cells9040848

Cited by 11 publications

(9 citation statements)

References 74 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Interestingly, patients who suffer from LSDs exhibit dysfunctional mitochondria, impairment of autophagy and accumulation of cytoplasmic protein aggregates [ 29 , 30 ]. Moreover, recently Coazzoli et al showed that another key enzyme in sphingolipid metabolism, acid sphingomyelinase, has an important role in the regulation of mitochondrial function and morphology in melanoma cells [ 31 ].…”

Section: Discussionmentioning

confidence: 99%

Ablation of Acid Ceramidase Impairs Autophagy and Mitochondria Activity in Melanoma Cells

Lai

Rocca

Amato

et al. 2021

IJMS

View full text Add to dashboard Cite

Cutaneous melanoma is often resistant to therapy due to its high plasticity, as well as its ability to metabolise chemotherapeutic drugs. Sphingolipid signalling plays a pivotal role in its progression and metastasis. One of the ways melanoma alters sphingolipid rheostat is via over-expression of lysosomal acid ceramidase (AC), which catalyses the hydrolysis of pro-apoptotic long-chain ceramides into sphingosine and fatty acid. In this report, we examine the role of acid ceramidase in maintaining cellular homeostasis through the regulation of autophagy and mitochondrial activity in melanoma cell lines. We show that under baseline conditions, wild-type melanoma cells had 3-fold higher levels of the autophagy marker, microtubule-associated proteins 1A/1B light chain 3B (LC3 II), compared to AC-null cells. This difference was further magnified after cell starvation. Moreover, we noticed autophagy impairment in A375 AC-null cells, possibly due to local accumulation of non-metabolized ceramides. Nonetheless, we observed that AC-null cells exhibited a significant increase in mitochondrial membrane potential compared to control cells. Consistent with this observation, we found that, after total starvation, ~30% of AC-null cells undergo apoptosis compared to ~6% of wild-type cells. As expected, AC transfection restored viability in A375 AC-null cells. Together, these findings suggest that AC-null melanoma cells change and adapt their metabolism to survive in the absence of AC, although in a way that does not allow them to cope with the stress of nutrient deprivation.

show abstract

Section: Discussionmentioning

confidence: 99%

Ablation of Acid Ceramidase Impairs Autophagy and Mitochondria Activity in Melanoma Cells

Lai

Rocca

Amato

et al. 2021

IJMS

View full text Add to dashboard Cite

show abstract

“…Purified RNA was then reverse transcribed into cDNA with MultiScribe Reverse Transcriptase, oligo(dT), and a random hexamer primer mix (all from Applied Biosystems Life Technologies, CA, USA). Quantitative evaluation of the target gene was then performed by using an ABI Prism 7500 sequence detection system (Applied Biosystems Life Technologies) with Power SYBR Green PCR Master Mix reagent and specific primers (mSREBP2: sense, 5′-gcgttctggagaccatgga-3′, antisense, 5′-acaaagttgctctgaaaacaaatca-3′; mGK [19]: sense, 5′-ccctgagtggcttacagttc-3′, antisense, 5′-acggatgtggagtgttgaagc-3′; mGLUT2 [19]: sense, 5′-cctcaagaggtaataatatccc-3′, antisense, 5′-ccatcaagagggctccagtc-3′; mHMGCR [19]: sense, 5′-cttgtggaatgccttgtgattg-3′, antisense, 5′-agccgaagcagcacatgat-3′; mHMGCS: sense, 5′-gccgtgaactgggtcgaa-3′, antisense, 5′-gcatatatagcaatgtctcctgcaa-3′; mPFKFB3 [20]: sense, 5′-ctatcccacgggagagtcc-3′, antisense, 5′-tggcgctctaattccatga-3′; mASMASE [21]: sense, 5′-tgggactcctttggatggg-3′, antisense, 5′-cggcgctatggcactgaat-3′; mG6PD [22]: sense, 5′-ccggaaactggctgtgcgct-3′, antisense, 5′-ccaggtcacccgatgcaccc-3′ and mSLC7A5/LAT1 [23]: sense, 5′-atatcacgctgctcaacggtg-3′, antisense, 5′-ctccagcatgtaggcgtagtc-3′). The PCR cycling conditions were 95 °C (15 s), 40 cycles at 95 °C (15 s), and 60 °C (1 min).…”

Section: Quantitative Real-time Pcr (Rt-qpcr)mentioning

confidence: 99%

The Nox2-ROS-Nlrp3 Inflammasome Signaling Stimulates in the Hematopoietic Stem/Progenitor Cells Lipogenesis to Facilitate Membrane Lipid Raft Formation

Abdelbaset‐Ismail

Ciechanowicz

Bujko

et al. 2022

Stem Cell Rev and Rep

View full text Add to dashboard Cite

Proliferation, metabolism, and migration of hematopoietic stem/progenitor cells (HSPCs) are coordinated by receptors expressed on outer cell membranes that are integrated into microdomains, known as membrane lipid rafts (MLRs). These structures float freely in the cell membrane bilayer and are enriched in cholesterol and sphingolipids for their functional integrity. Receptors, if expressed in MLRs, have prolonged occupancy on the cell surface and enhanced signaling power. Based on this, we have become interested in the regulation of synthesis of MLRs components in HSPCs. To address this, we tested the effect of selected factors that promote proliferation or migration and their potential involvement in the synthesis of MLRs components in HSPCs. Based on our previous research showing that HSPCs from Nox2-KO and Nlrp3-KO mice display a profound defect in MLRs formation, we focused on the role of Nox2-ROS-Nlrp3 inflammasome in regulating lipogenesis in HSPCs. We found that while at steady state conditions, Nox2-derived ROS is required for a proper expression of enzymes regulating lipogenesis, during inflammation, this effect is augmented by Nlrp3 inflammasome. Thus, our data sheds new light on the regulation of lipogenesis in HSPCs and the involvement of the Nox2-ROS-Nlrp3 inflammasome axis that differently regulates lipogenesis at steady state conditions and in response to inflammation, modulating MLRs-mediated responsiveness of these cells to external stimuli. Graphical Abstract

show abstract

“…In B16 cells, chloroquine triggered lysosomal accumulation and oxidative stress, resulting in mitochondrial depolarization and apoptotic/necrotic cell death. In this respect, chloroquine treatment of cells resistant to cisplatin, characterised also by an alteration of oxidative metabolism, led to the re-sensitization to the chemotherapeutic drug through the modulation of mammalian target of rapamycin (mTOR)-mediated autophagy [ 82 , 83 ]. A similar scenario was observed in calorie-restricted mice in which chloroquine reduced melanoma growth and necrosis of tumour cells.…”

Section: Melanoma Cell Death and Survival: Simultaneous Regulation Of Oxidative Stress System And Autophagy Machinerymentioning

confidence: 99%

Oxidative Stress and Autophagy as Key Targets in Melanoma Cell Fate

Catalani

Giovarelli

Zecchini

et al. 2021

Cancers

Self Cite

View full text Add to dashboard Cite

Melanoma originates from the malignant transformation of melanocytes and is one of the most aggressive forms of cancer. The recent approval of several drugs has increased the chance of survival although a significant subset of patients with metastatic melanoma do not show a long-lasting response to these treatments. The complex cross-talk between oxidative stress and the catabolic process autophagy seems to play a central role in all aspects of melanoma pathophysiology, from initiation to progression and metastasis, including drug resistance. However, determining the fine role of autophagy in cancer death and in response to redox disruption is still a fundamental challenge in order to advance both basic and translational aspects of this field. In order to summarize the interactions among reactive oxygen and nitrogen species, autophagy machinery and proliferation/growth/death/apoptosis/survival, we provide here a narrative review of the preclinical evidence for drugs/treatments that modulate oxidative stress and autophagy in melanoma cells. The significance and the potential for pharmacological targeting (also through multiple and combination approaches) of these two different events, which can contribute independently or simultaneously to the fate of melanoma, may help to define new processes and their interconnections underlying skin cancer biology and unravel new reliable approaches.

show abstract

Acid Sphingomyelinase Downregulation Enhances Mitochondrial Fusion and Promotes Oxidative Metabolism in a Mouse Model of Melanoma

Cited by 11 publications

References 74 publications

Ablation of Acid Ceramidase Impairs Autophagy and Mitochondria Activity in Melanoma Cells

Ablation of Acid Ceramidase Impairs Autophagy and Mitochondria Activity in Melanoma Cells

The Nox2-ROS-Nlrp3 Inflammasome Signaling Stimulates in the Hematopoietic Stem/Progenitor Cells Lipogenesis to Facilitate Membrane Lipid Raft Formation

Oxidative Stress and Autophagy as Key Targets in Melanoma Cell Fate

Contact Info

Product

Resources

About