Publications

@article{Cardenas2024-ev,

title = {NANOG controls testicular germ cell tumour stemness through  regulation of MIR9-2},

author = {Ryan P Cardenas and Ahmad Zyoud and Alan McIntyre and Ramiro Alberio and Nigel P Mongan and Cinzia Allegrucci},

doi = {10.1186/s13287-024-03724-1},

year  = {2024},

date = {2024-05-01},

urldate = {2024-05-01},

journal = {Stem Cell Res. Ther.},

volume = {15},

number = {1},

pages = {128},

publisher = {Springer Science and Business Media LLC},

abstract = {BACKGROUND: Testicular germ cell tumours (TGCTs) represent a 

 clinical challenge; they are most prevalent in young individuals 

 and are triggered by molecular mechanisms that are not fully 

 understood. The origin of TGCTs can be traced back to primordial 

 germ cells that fail to mature during embryonic development. 

 These cells express high levels of pluripotency factors, 

 including the transcription factor NANOG which is highly 

 expressed in TGCTs. Gain or amplification of the NANOG locus is 

 common in advanced tumours, suggesting a key role for this 

 master regulator of pluripotency in TGCT stemness and 

 malignancy. METHODS: In this study, we analysed the expression 

 of microRNAs (miRNAs) that are regulated by NANOG in TGCTs via 

 integrated bioinformatic analyses of data from The Cancer Genome 

 Atlas and NANOG chromatin immunoprecipitation in human embryonic 

 stem cells. Through gain-of-function experiments, MIR9-2 was 

 further investigated as a novel tumour suppressor regulated by 

 NANOG. After transfection with MIR9-2 mimics, TGCT cells were 

 analysed for cell proliferation, invasion, sensitivity to 

 cisplatin, and gene expression signatures by RNA sequencing. 

 RESULTS: For the first time, we identified 86 miRNAs regulated 

 by NANOG in TGCTs. Among these, 37 miRNAs were differentially 

 expressed in NANOG-high tumours, and they clustered TGCTs 

 according to their subtypes. Binding of NANOG within 2 kb 

 upstream of the MIR9-2 locus was associated with a negative 

 regulation. Low expression of MIR9-2 was associated with tumour 

 progression and MIR9-2-5p was found to play a role in the 

 control of tumour stemness. A gain of function of MIR9-2-5p was 

 associated with reduced proliferation, invasion, and sensitivity 

 to cisplatin in both embryonal carcinoma and seminoma tumours. 

 MIR9-2-5p expression in TGCT cells significantly reduced the 

 expression of genes regulating pluripotency and cell division, 

 consistent with its functional effect on reducing cancer 

 stemness. CONCLUSIONS: This study provides new molecular 

 insights into the role of NANOG as a key determinant of 

 pluripotency in TGCTs through the regulation of MIR9-2-5p, a 

 novel epigenetic modulator of cancer stemness. Our data also 

 highlight the potential negative feedback mediated by MIR9-2-5p 

 on NANOG expression, which could be exploited as a therapeutic 

 strategy for the treatment of TGCTs.},

keywords = {Differentiation, MIR-9, NANOG, Pluripotency, stemness, Testicular germ cell tumours},

pubstate = {published},

tppubtype = {article}

}

@article{Martinez-Espuga2023-gt,

title = {Intestinal cell differentiation and phenotype in 2D and 3D 

 cell culture models},

author = {Magda Mart\'{i}nez-Espuga and Alvaro Mata and Paloma Ord\'{o}\~{n}ez-Mor\'{a}n},

year  = {2023},

date = {2023-01-01},

journal = {Methods Mol. Biol.},

volume = {2650},

pages = {235\textendash243},

abstract = {Three-dimensional (3D) culture models are more physiologically 

 relevant than two-dimensional (2D) cell culture models. 2D 

 approaches cannot reproduce the complexity of the tumor 

 microenvironment and are less able to translate biological 

 insights; and drug response studies have many limitations to be 

 extrapolated to the clinics. Here, we use the Caco-2 colon cancer 

 cell line, which is an immortalized human epithelial cell line 

 that under specific conditions can polarize and differentiate 

 into a villus-like phenotype. We describe cell differentiation 

 and cell growth in both 2D and 3D culture conditions, concluding 

 that cell morphology, polarity, proliferation and differentiation 

 are highly dependent on the type of cell culture system.},

keywords = {2D, 3D, Caco-2 cells, Differentiation, Gene expression, Markers},

pubstate = {published},

tppubtype = {article}

}

@article{Bonilla-Diaz2023-eh,

title = {Differentiated epithelial cells of the gut},

author = {Andrea Bonilla-D\'{i}az and Paloma Ord\'{o}\~{n}ez-Mor\'{a}n},

year  = {2023},

date = {2023-01-01},

journal = {Methods Mol. Biol.},

volume = {2650},

pages = {3\textendash16},

abstract = {The intestine is a prime example of self-renewal where stem cells 

 give rise to progenitor cells called transit-amplifying cells 

 which differentiate into more specialized cells. There are two 

 intestinal lineages: the absorptive (enterocytes and microfold 

 cells) and the secretory (Paneth cells, enteroendocrine, goblet 

 cells, and tuft cells). Each of these differentiated cell types 

 has a role in creating an ``ecosystem'' to maintain intestinal 

 homeostasis. Here, we summarize the main roles of each cell type.},

keywords = {Colon, Differentiation, Enterocytes, Enteroendocrine cells, Goblet cells, Intestine, Microfold cells, Paneth cells, Tuft   cells},

pubstate = {published},

tppubtype = {article}

}

@article{Gajera2023-kn,

title = {In vitro and in vivo assays for testing retinoids effect on 

 intestinal progenitors' lineage commitments},

author = {Krishna R Gajera and Kathryn L Fair and Gordon W Moran and Nicholas R F Hannan and Joerg Huelsken and Paloma Ord\'{o}\~{n}ez-Mor\'{a}n},

year  = {2023},

date = {2023-01-01},

journal = {Methods Mol. Biol.},

volume = {2650},

pages = {53\textendash61},

abstract = {The intestine consists of epithelial cells surrounded by a 

 complex environment as mesenchymal cells and the gut microbiota. 

 With its impressive stem cell regeneration capability, the 

 intestine is able to constantly replenish cells lost through 

 apoptosis or abrasion by food passing through. Over the past 

 decade, researchers have identified signaling pathways involved 

 in stem cell homeostasis such as retinoids pathway. Retinoids are 

 also involved in cell differentiation of healthy and cancer 

 cells. In this study, we describe several approaches in vitro and 

 in vivo to further investigate the effect of retinoids on stem 

 cells, progenitors, and differentiated intestinal cells.},

keywords = {Absorptive, Differentiation, Intestine, Organoids, Retinoids, Secretory, Stem cells},

pubstate = {published},

tppubtype = {article}

}

@article{Marva2023-zw,

title = {Novel approach to measure transepithelial electrical resistance 

 in intestinal cells},

author = {Gurveer Marva and Seyda \"{U}nsal and Andrew V Benest and David O Bates and Paloma Ord\'{o}\~{n}ez-Mor\'{a}n},

year  = {2023},

date = {2023-01-01},

journal = {Methods Mol. Biol.},

volume = {2650},

pages = {35\textendash42},

abstract = {The technique electric cell-substrate impedance sensing (ECIS) 

 can be used to detect and monitor the behavior of intestinal 

 cells. The methodology presented was designed to achieve results 

 within a short time frame, and it was tailored to use a colonic 

 cancer cell line. Differentiation of intestinal cancer cells has 

 previously been reported to be regulated by retinoic acid (RA). 

 Here, colonic cancer cells were cultured in the ECIS array before 

 being treated with RA, and any changes in response to RA were 

 monitored after treatment. The ECIS recorded changes in impedance 

 in response to the treatment and vehicle. This methodology poses 

 as a novel way to record the behavior of colonic cells and opens 

 new avenues for in vitro research.},

keywords = {Caco-2 cells, Colon, Differentiation, ECIS, Intestine, TEER},

pubstate = {published},

tppubtype = {article}

}

@article{Mahmoud2022-ls,

title = {Challenges for Triple Negative Breast Cancer treatment: 

 Defeating heterogeneity and cancer stemness},

author = {Rinad Mahmoud and Paloma Ord\'{o}\~{n}ez-Mor\'{a}n and Cinzia Allegrucci},

year  = {2022},

date = {2022-09-01},

journal = {Cancers (Basel)},

volume = {14},

number = {17},

pages = {4280},

publisher = {MDPI AG},

abstract = {The Triple Negative Breast Cancer (TNBC) subtype is known to 

 have a more aggressive clinical course compared to other breast 

 cancer subtypes. Targeted therapies for this type of breast 

 cancer are limited and patients are mostly treated with 

 conventional chemo- and radio-therapies which are not specific 

 and do not target resistant cells. Therefore, one of the major 

 clinical challenges is to find compounds that target the 

 drug-resistant cell populations which are responsible for 

 reforming secondary tumours. The molecular profiling of the 

 different TNBC subtypes holds a promise for better defining 

 these resistant cells specific to each tumour. To this end, a 

 better understanding of TNBC heterogeneity and cancer stemness 

 is required, and extensive genomic analysis can help to 

 understand the disease complexity and distinguish new molecular 

 drivers that can be targeted in the clinics. The use of 

 persister cancer cell-targeting therapies combined with other 

 therapies may provide a big advance to improve TNBC patients' 

 survival.},

keywords = {Differentiation, drug resistance, persister cells, phenotype, stemness, triple negative breast cancer, tumour heterogeneity},

pubstate = {published},

tppubtype = {article}

}

@article{Ahmed2019-eb,

title = {Repurposing antibacterial AM404 as a potential anticancer drug 

 for targeting colorectal cancer stem-like cells},

author = {Mehreen Ahmed and Nicholas Jinks and Roya Babaei-Jadidi and Hossein Kashfi and Marcos Castellanos-Uribe and Sean T May and Abhik Mukherjee and Abdolrahman S Nateri},

year  = {2019},

date = {2019-12-01},

journal = {Cancers (Basel)},

volume = {12},

number = {1},

pages = {106},

publisher = {MDPI AG},

abstract = {Tumour-promoting inflammation is involved in colorectal cancer 

 (CRC) development and therapeutic resistance. However, the 

 antibiotics and antibacterial drugs and signalling that regulate 

 the potency of anticancer treatment upon forced differentiation 

 of cancer stem-like cell (CSC) are not fully defined yet. We 

 screened an NIH-clinical collection of the small-molecule 

 compound library of antibacterial/anti-inflammatory agents that 

 identified potential candidate drugs targeting CRC-SC for 

 differentiation. Selected compounds were validated in both in 

 vitro organoids and ex vivo colon explant models for their 

 differentiation induction, impediment on neoplastic cell growth, 

 and to elucidate the mechanism of their anticancer activity. We 

 initially focused on AM404, an anandamide uptake inhibitor. 

 AM404 is a metabolite of acetaminophen with antibacterial 

 activity, which showed high potential in preventing CRC-SC 

 features, such as stemness/de-differentiation, migration and 

 drug-resistance. Furthermore, AM404 suppressed the expression of 

 FBXL5 E3-ligase, where AM404 sensitivity was mimicked by 

 FBXL5-knockout. This study uncovers a new molecular mechanism 

 for AM404-altering FBXL5 oncogene which mediates 

 chemo-resistance and CRC invasion, thereby proposes to repurpose 

 antibacterial AM404 as an anticancer agent.},

keywords = {AM404, cancer stem cells, colonosphere, CRC, Differentiation, drug screening, FBXL5 E3-ligase, patient derived organoids, resistance and metastasis, tissue explants},

pubstate = {published},

tppubtype = {article}

}