HDAC6 inhibition decreases leukemic stem cell expansion driven by Hedgehog hyperactivation by restoring primary ciliogenesis

Pezzotta, Alex; Gentile, Ilaria; Genovese, Donatella; Totaro, Maria Grazia; Battaglia, Cristina; Leung, Anskar Y. H.; Fumagalli, Monica; Parma, Matteo; Cazzaniga, G; Fazio, Grazia; Alcalay, Myriam; Marozzi, Anna; Pistocchi, Anna

doi:10.1016/j.phrs.2022.106378

Cited by 5 publications

(17 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To determine if the effect of the simultaneous Hh /HDAC6 inhibition on the autophagic process are specifically due to these pathways, we used our previously generated zebrafish model in conjunction with hyperactivation of the Hh signaling. Indeed, we demonstrated the existence of a crosstalk between these two signaling pathways, as the overexpression of the Hh ligand shh also elicited the upregulation of hdac6 [ 21 ]. Using this in vivo model, we verified that the number of Lc3b autophagosomes was significantly decreased in the shh -mRNA-injected embryos from the Tg( CMV:eGFPmap1Lc3b ) line as compared to the control embryos treated with the DMSO ( Figure 6 A–C).…”

Section: Resultsmentioning

confidence: 99%

“…Zebrafish shh -mRNA was in vitro transcribed as previously described [ 21 ]. In this process, 100 pg/embryo of the shh -mRNA was injected into one-cell-stage of the Tg( CMV:eGFP-map1Lc3b ) embryos together with the phenol red vital dye as a tracer of microinjection efficacy.…”

Section: Methodsmentioning

confidence: 99%

“…Both HDAC6 and Hh have a prominent role in the regulation of autophagy at multiple levels [ 19 , 20 ]; therefore, the effects of their combined inhibition on this process could be an intriguing feature to examine. Interestingly, we recently demonstrated that there is crosstalk between Hh and HDAC6 and that their inhibition effectively reduces the expansion of hematopoietic stem progenitor cells presented in zebrafish models of myeloid leukemia [ 21 ]. Zebrafish, ( Danio rerio ), is a well-established animal model used to study human diseases, including cancer.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Combined Inhibition of Hedgehog and HDAC6: In Vitro and In Vivo Studies Reveal a New Role for Lysosomal Stress in Reducing Glioblastoma Cell Viability

Pezzotta

Brioschi

Carbone

et al. 2023

IJMS

Self Cite

View full text Add to dashboard Cite

Glioblastoma multiforme (GBM) is the most common and malignant brain tumor in adults. The invasiveness and the rapid progression that characterize GBM negatively impact patients’ survival. Temozolomide (TMZ) is currently considered the first-choice chemotherapeutic agent. Unfortunately, over 50% of patients with GBM do not respond to TMZ treatment, and the mutation-prone nature of GBM enables the development of resistance mechanisms. Therefore, efforts have been devoted to the dissection of aberrant pathways involved in GBM insurgence and resistance in order to identify new therapeutic targets. Among them, sphingolipid signaling, Hedgehog (Hh) pathway, and the histone deacetylase 6 (HDAC6) activity are frequently dysregulated and may represent key targets to counteract GBM progression. Given the positive correlation between Hh/HDAC6/sphingolipid metabolism in GBM, we decided to perform a dual pharmacological inhibition of Hh and HDAC6 through cyclopamine and tubastatin A, respectively, in a human GMB cell line and zebrafish embryos. The combined administration of these compounds elicited a more significant reduction of GMB cell viability than did single treatments in vitro and in cells orthotopically transplanted in the zebrafish hindbrain ventricle. We demonstrated, for the first time, that the inhibition of these pathways induces lysosomal stress which results in an impaired fusion of lysosomes with autophagosomes and a block of sphingolipid degradation in GBM cell lines. This condition, which we also recapitulated in zebrafish embryos, suggests an impairment of lysosome-dependent processes involving autophagy and sphingolipid homeostasis and might be instrumental in the reduction of GBM progression.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Combined Inhibition of Hedgehog and HDAC6: In Vitro and In Vivo Studies Reveal a New Role for Lysosomal Stress in Reducing Glioblastoma Cell Viability

Pezzotta

Brioschi

Carbone

et al. 2023

IJMS

Self Cite

View full text Add to dashboard Cite

show abstract

“…Embryos dissociation was obtained 88 . GFP+ cells were sorted from Tg( fli1a: GFP) y1 63 embryos at 32-34 hpf using a BD FACSMelody Cell Sorter (BD Bioscience, Franklin Lakes, NJ, USA).…”

Section: Methodsmentioning

confidence: 99%

ADA2 regulates inflammation and hematopoietic stem cell emergence via the A2bR pathway in zebrafish

Brix,

Belleri,

Pezzotta

et al. 2024

Commun Biol

Self Cite

View full text Add to dashboard Cite

Deficiency of adenosine deaminase 2 (DADA2) is an inborn error of immunity caused by loss-of-function mutations in the adenosine deaminase 2 (ADA2) gene. Clinical manifestations of DADA2 include vasculopathy and immuno-hematological abnormalities, culminating in bone marrow failure. A major gap exists in our knowledge of the regulatory functions of ADA2 during inflammation and hematopoiesis, mainly due to the absence of an ADA2 orthologue in rodents. Exploring these mechanisms is essential for understanding disease pathology and developing new treatments. Zebrafish possess two ADA2 orthologues, cecr1a and cecr1b, with the latter showing functional conservation with human ADA2. We establish a cecr1b-loss-of-function zebrafish model that recapitulates the immuno-hematological and vascular manifestations observed in humans. Loss of Cecr1b disrupts hematopoietic stem cell specification, resulting in defective hematopoiesis. This defect is caused by induced inflammation in the vascular endothelium. Blocking inflammation, pharmacological modulation of the A2r pathway, or the administration of the recombinant human ADA2 corrects these defects, providing insights into the mechanistic link between ADA2 deficiency, inflammation and immuno-hematological abnormalities. Our findings open up potential therapeutic avenues for DADA2 patients.

show abstract

“…Aberrant hedgehog (Hh) signaling is also involved in the maintenance of CSCs, which is activated by the binding with Hh ligands, such as sonic hedgehog (Shh), to their receptors Ptch1 or Ptch2 (Yu et al, 2022). CSCs respond to Shh‐activated Hh signaling by maintaining a stemness signature through the influences on pluripotency genes, including Sox2, Nanog, and so on (Pezzotta et al, 2022). In our previous work, we found that Oct4‐driven reprogramming of astrocytes into induced neural stem cells (iNSCs) was potentiated with continuous Shh stimulation, and this event is mediated by the crosstalk between Sox2 /Shh‐targeted downstream signals and PI3K/Cdk2/Smurf2 signaling (H. Yang et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

Dauricine attenuates Oct4/sonic hedgehog co‐activated stemness and induces reactive oxygen species‐mediated mitochondrial apoptosis via AKT/β‐catenin signaling in human neuroblastoma and glioblastoma stem‐like cells

Liu,

Yang,

Cheng

et al. 2023

Phytotherapy Research

View full text Add to dashboard Cite

Neuroblastoma and glioblastoma are primary malignant tumors of the nervous system, with frequent relapse and limited clinical therapeutic drugs. The failure of their treatment is due to the tumor cells exhibiting cancer stem‐like cells (CSLCs) properties. Octamer binding transcription factor 4 (Oct4) is involved in mediating CSLCs, our previous work found that Oct4‐driven reprogramming of astrocytes into induced neural stem cells was potentiated with continuous sonic hedgehog (Shh) stimulation. In this study, we aimed to study the importance of Oct4 and Shh combination in the stemness properties induction of neuroblastoma and glioblastoma cells, and evaluate the anti‐stemness effect of dauricine (DAU), a natural product of bis‐benzylisoquinoline alkaloid. The effect of Oct4 and Shh co‐activation on cancer stemness was evaluated by tumor spheres formation model and flow cytometry analysis. Then the effects of DAU on SH‐SY5Y and T98‐G cells were assessed by the MTT, colony formation, and tumor spheres formation model. DAU acts on Oct4 were verified using the Western blotting, MTT, and so on. Mechanistic studies were explored by siRNA transfection assay, Western blotting, and flow cytometry analysis. We identified that Shh effectively improved Oct4‐mediated generation of stemness in SH‐SY5Y and T98‐G cells, and Oct4 and Shh co‐activation promoted cell growth, the resistance of apoptosis. In addition, DAU, a natural product, was found to be able to attenuate Oct4/Shh co‐activated stemness and induce cell cycle arrest and apoptosis via blocking AKT/β‐catenin signaling in neuroblastoma and glioblastoma, which contributed to the neuroblastoma and glioblastoma cells growth inhibition by DAU. In summary, our results indicated that the treatment of DAU may be served as a potential therapeutic method in neuroblastoma and glioblastoma.

show abstract

HDAC6 inhibition decreases leukemic stem cell expansion driven by Hedgehog hyperactivation by restoring primary ciliogenesis

Cited by 5 publications

References 37 publications

Combined Inhibition of Hedgehog and HDAC6: In Vitro and In Vivo Studies Reveal a New Role for Lysosomal Stress in Reducing Glioblastoma Cell Viability

Combined Inhibition of Hedgehog and HDAC6: In Vitro and In Vivo Studies Reveal a New Role for Lysosomal Stress in Reducing Glioblastoma Cell Viability

ADA2 regulates inflammation and hematopoietic stem cell emergence via the A2bR pathway in zebrafish

Dauricine attenuates Oct4/sonic hedgehog co‐activated stemness and induces reactive oxygen species‐mediated mitochondrial apoptosis via AKT/β‐catenin signaling in human neuroblastoma and glioblastoma stem‐like cells

Contact Info

Product

Resources

About