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Biology subjects

Marcato, P.

Publications and source records attributed to Marcato, P..

3 recordsLinked to original sources

CLM296: a highly selective inhibitor targeting ALDH1A3-driven tumor growth and metastasis in breast cancer

Aldehyde dehydrogenase 1A3 (ALDH1A3) increases tumor growth, metastasis, and chemoresistance in many solid tumors, including triple-negative breast cancer (TNBC), glioblastoma, melanoma, lung, and colon cancers, yet no clinically approved inhibitors exist. Here, we present CLM296, a novel and highly selective ALDH1A3 inhibitor designed to address this unmet need. CLM296 exhibits potent inhibition of ALDH1A3 activity in TNBC cells (half-maximal inhibitory concentration = 2 nM) with no off-target effects on the highly homologous ALDH1A1 isoform. RNA sequencing confirmed its specificity, demonstrating selective suppression of ALDH1A3-regulated gene expression only, and a lack of effect in control cells that have minimal ALDH1A3 expression. Transwell assays showed that CLM296 reduced the increased invasion of cells induced by ALDH1A3. Once daily dosing of 4mg/kg CLM296 in mice specifically reduced ALDH1A3-mediated gene expression in tumors and impeded ALDH1A3-driven tumor growth and lung metastasis in TNBC xenografts. There was no observed toxicity in the mice as evidenced by stable mouse body weights and no significant changes in blood creatinine and ALT levels. Pharmacokinetic studies of CLM296 revealed broad tissue distribution, including tumor, lung, liver, and brain. With oral administration the terminal elimination half-life of CLM296 exceeded 12 hours, resulting in sustained ALDH1A3-inhibiting concentrations beyond 24 hours. Together, these findings establish CLM296 as a potential first-in-class ALDH1A3 inhibitor with high selectivity for ALDH1A3, favorable pharmacokinetics, and a positive preclinical safety profile. CLM296 represents a promising therapeutic candidate to complement standard-of-care treatments in ALDH1A3+ cancers.

cancer biology↗

In Vitro Inhibition of SARS-CoV-2 Infection by Bromhexine hydrochloride

The world is enduring the SARS CoV-2 pandemic, and although extensive research has been conducted on the issue, only a few antivirals have been approved to treat patients with COVID-19. Bromhexine hydrochloride was previously identified as a potent inhibitor of TMPRSS2, an essential protease for ACE-2 virus receptor interactions. In the present study, we investigated whether bromhexine treatment could reduce SARS CoV-2 replication in vitro. To evaluate bromhexines effectiveness against SARS COV-2 infection, viral load was measured using Caco-2 cell lines expressing TMPRSS2. Our molecular docking results indicate that bromhexine displays an affinity with the active site of TMPRSS2. The drug significantly inhibited SARS CoV-2, both parental and P1 variant strains, infection in the Caco-2 cell line, reducing about 40% of SARS-CoV-2 entrance and about 90% of viral progeny in the supernatant 48h post-infection. Furthermore, bromhexine did not exhibit any direct virucidal activity on SARS CoV-2. In conclusion, bromhexine hydrochloride efficiently disrupts SARS CoV-2 infection in vitro and has the potential to become an effective antiviral agent in COVID-19 treatment.

pharmacology and toxicology↗

Human-specific GAPDH RT-qPCR is an accurate and sensitive method of xenograft metastasis quantification

Metastasis is the primary cause of cancer-related mortality. Having experimental models that accurately reflect changes in the metastatic burden is imperative for developing improved treatments and a better understanding of the disease. The murine xenograft tumor model mimics the human scenario and provides a platform for in vivo and ex vivo metastasis quantification analyses. Histological analysis of hematoxylin and eosin (H&E) stained thin sections has been the gold standard for quantifying metastasis ex vivo but gaining favor for its ease and accuracy is reverse transcription-qualitative polymerase chain reaction (RT-qPCR). Herein we directly compare histological and RT-qPCR-based methods for quantifying lung metastasis in a murine xenograft tumor model. Furthermore, we have introduced a variation of the RT-qPCR method; human-specific glyceraldehyde 3-phosphate dehydrogenase (GAPDH) RT-qPCR, which allows quantification of metastasis in xenograft models, without the requirement of overexpression of exogenous genes. Human-specific GAPDH RT-qPCR detected increased lung metastasis resulting from aldehyde dehydrogenase 1A3 (ALDH1A3) expression in MDA-MB-231 breast cancer cells orthotopically implanted in NOD/SCID mice. Further, in the xenograft tumor model, human-specific GAPDH RT-qPCR was more sensitive and cost-effective than quantification of lung metastasis by histological analysis of H&E stained fixed thin sections. The two assays were highly correlative in terms of determining relative metastatic burden, suggesting that the human-specific GAPDH RT-qPCR method could be used as a standard method for quantification of disseminated human cells in murine xenograft models.

cancer biology↗