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CRISPR/Cas system: An emerging technology in stem cell research

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World Journal of Stem Cells 2019年第11期11卷 937-956页

作者： Maria Teresa Valenti michela Serena Luca Dalle Carbonare Donato Zipeto Department of Medicine Section of Internal Medicine DUniversity of VeronaVerona 37134Italy Department of Biochemistry University of OxfordOxford OX13QUUnited Kingdom Department of Neurosciences Biomedicine and Movement SciencesLaboratory of Molecular BiologyVerona 37134Italy

The identification of new and even more precise technologies for modifying and manipulating the genome has been a challenge since the discovery of the DNA double *** ability to modify selectively specific genes provides a powerful tool for characterizing gene functions,performing gene therapy,correcting specific genetic mutations,eradicating diseases,engineering cells and organisms to achieve new and different functions and obtaining transgenic animals as models for studying specific *** regularly interspaced short palindromic repeats(CRISPR)/Cas9 technology has recently revolutionized genome *** application of this new technology to stem cell research allows disease models to be developed to explore new therapeutic *** possibility of translating new systems of molecular knowledge to clinical research is particularly appealing for addressing degenerative *** this review,we describe several applications of CRISPR/Cas9 to stem cells related to degenerative *** addition,we address the challenges and future perspectives regarding the use of CRISPR/Cas9 as an important technology in the medical sciences.

关键词： Gene editing CRISPR/Cas9 Stem cells Degenerative diseases

Human induced pluripotent stem cells for monogenic disease modelling and therapy

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World Journal of Stem Cells 2016年第4期8卷 118-135页

作者： Paola Spitalieri Valentina Rosa Talarico michela Murdocca Giuseppe Novelli Federica Sangiuolo Department of Biomedicine and Prevention Tor Vergata University

Recent and advanced protocols are now available to derive human induced pluripotent stem cells (hiPSCs) from patients affected by genetic diseases. No curative treatments are available for many of these diseases; thus, hiPSCs represent a major impact on patient’ health. hiPSCs represent a valid model for the in vitro study of monogenic diseases, together with a better comprehension of the pathogenic mechanisms of the pathology, for both cell and gene therapy protocol applications. Moreover, these pluripotent cells represent a good opportunity to test innovative pharmacological treatments focused on evaluating the efficacy and toxicity of novel drugs. Today, innovative gene therapy protocols, especially gene editing-based, are being developed, allowing the use of these cells not only as in vitro disease models but also as an unlimited source of cells useful for tissue regeneration and regenerative medicine, eluding ethical and immune rejection problems. In this review, we will provide an up-to-date of modelling monogenic disease by using hiPSCs and the ultimate applications of these in vitro models for cell therapy. We consider and summarize some peculiar aspects such as the type of parental cells used for reprogramming, the methods currently used to induce the transcription of the reprogramming factors, and the type of iPSC-derived differentiated cells, relating them to the genetic basis of diseases and to their inheritance model.

关键词： Human induced pluripotent stem cells Gene therapy Monogenic diseases Gene editing Foetal cells Reprogramming techniques Differentiation

Eye reflux:an ocular extraesophageal manifestation of gastric reflux

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International Journal of Ophthalmology(English edition) 2018年第9期11卷 1503-1507页

作者： Danilo Mazzacane Valerio Damiani michela Silvestri Giorgio Ciprandi Pierfranco Marino Asst Pavia and Asst Melegnano and Martesana Pavia 27100Italy DMG Medical Department Pomezia 00071Italy Istituto Giannina Gaslini Genoa 16147Italy Ospedale Policlinico San Martino Genoa 16132Italy Dipartimento di Scienze della Salute Universita del MoliseCampobasso 86100Italy

AIM： To suspect laryngopharyngeal reflux（LPR） in patients with ocular surface disease（OSD）. METHODS： The present study evaluated a group of subjects with OSD assessing the Ocular Surface Disease Index（OSDI） and the Reflux Symptom Index（RSI） to detect patients with suspected LPR and define a possible relationship between ***： Two hundred and ninety subjects（175 females, mean age： 60.41±15.68y） were consecutively visited at ophthalmologist offices. One hundred and one（34%） patients had pathological RSI（〉13） and consequently a suspected ***： The current study shows that suspected LPR may be common（34%） in patients with OSD and a suspected LPR may be considered in OSD patients when RSI score is 〉13 and OSDI score is 〉42.

关键词： gastric reflux eye reflux ocular surface disease index reflux symptom score

Shaping the future of pediatric liver health:unraveling the impact of the new metabolic-associated fatty liver disease definition

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Hepatobiliary Surgery and Nutrition 2023年第4期12卷 611-615页

作者： Stefano Ciardullo michela Vergani Celeste Ronchetti Gianluca Perseghin Department of Medicine and Surgery University of Milano BicoccaMilanItaly Department of Medicine and Rehabilitation Policlinico di MonzaMonzaItaly

Liver steatosis(i.e.,excessive triglyceride accumulation within the hepatocyte)is a very common finding in both the adult and pediatric *** the latter,epidemiologic data using validated diagnostic techniques(either liver biopsy or ultrasound-based technologies)have shown an alarmingly high prevalence,paralleling the ever-increasing rates of overweight and obesity.

关键词： Nonalcoholic fatty liver disease(NAFLD) metabolic-associated fatty liver disease(MAFLD) pediatrics definition fatty liver

18,X,Y aneuploidies and transmission electron microscopy studies in spermatozoa from five carriers of different reciprocal translocations

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Asian Journal of Andrology 2009年第3期11卷 325-332页

作者： Elena Moretti Nicola Antonio Pascarelli Valentina Giannerini michela Geminiani Cecilia Anichini Giulia Collodel Department of Biomedical Sciences Applied Biology Section University of Siena Siena 53100 Italy Department of Pediatrics Obstetrics and Reproductive Medicine University of Siena Siena 53100 Italy

We analysed ejaculated spermatozoa from five infertile men with different balanced reciprocal translocations to contribute to the study of meiotic segregation of chromosomes 18, X and Y and also to evaluate sperm morphology by transmission electron microscopy （TEM） analysis. Conventional lymphocyte karyotype analyses highlighted dif- ferent reciprocal balanced translocations： t（12; 13）, t（4;9）, t（X;8）, t（8; 10） and t（3; 16）. Semen analysis was performed by light and TEM. Fluorescence in situ hybridization was performed directly on sperm nuclei using centromeric probes for chromosomes 18, X and Y. The carriers of the balanced reciprocal translocations considered in the present study showed a very similar pattern of sperm pathologies： diffused presence of apoptosis and immaturity. All patients showed meiotic segregation derangements, highlighted by the presence of sperm diploidies and sex chromosome disomies particularly related to the failure of the first meiotic division. However, an increased incidence of chromosome 18 aneuploidy was detected in spermatozoa from t（X;8） and t（8;10） carriers. We have also reported values from sex chromosomes such as t（X;8）, although the X chromosome was involved in translocation. Since patients with reciprocal translocations and spermatogenetic impairment are candidates for intracytoplasmic sperm injection cycles, the study of sperm parameters, and particularly of the level of aneuploidy rates, would provide better information for couples at risk and would contribute to the data in the literature for a better understanding of the effects of chromosomal rearrangement on the whole meiotic process and, in particular, on chromosomes not involved in translocation.

关键词： electron microscopy fluorescence in situ hybridization altered karyotype spermatozoa reciprocal translocation

Anal metastasis from recurrent breast lobular carcinoma:A case report

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World Journal of Gastroenterology 2009年第11期15卷 1388-1390页

作者： Maria Puglisi Emanuela Varaldo michela Assalino Gianluca Ansaldo Giancarlo Torre Giacomo Borgonovo University of Genova Largo Benzi8.16132 GenovaItaly

We report a case of isolated gastrointestinal metastasis from breast Iobular carcinoma, which mimicked primary anal cancer. In July 2000, an 88-year-old woman presented with infiltrating Iobular cancer （pT1/G2/N2）. The patient received postoperative radiotherapy and hormonal therapy. Four years later, she presented with an anal polypoid lesion. The mass was removed for biopsy. Immunohistochemical staining suggested a breast origin. Radiotherapy was chosen for this patient, which resulted in complete regression of the lesion. The patient died 3 years after the first manifestation of gastrointestinal metastasis. According to the current literature, we consider the immunohistochemistry features that are essential to support the suspicion of gastrointestinal breast metastasis, and since we consider the gastrointestinal involvement as a sign of systemic disease, the therapy should be less aggressive and systemic.

关键词： Anal cancer Breast cancer InfiltratingIobular carcinoma Estrogen receptors Progesteronereceptors

FISSION1A, an Arabidopsis Tail-Anchored Protein, Is Localized to Three Subcellular Compartments

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Molecular Plant 2014年第8期7卷 1393-1396页

作者： Cristina Ruberti Alex Costa Emanuela Pedrazzini Fiorella Lo Schiavo michela Zottini Department of Biology University of Padua Via U. Bassi 58/B I-35131 Padua Italy MSU-DOE Plant Research Laboratory and Department of Plant Biology Michigan State University 612 WilsonRoad East Lansing MI 48824 USA Department of Biosciences University of Milan Via G. Celoria 26 I-20133 Milan Italy Institute of Biophysics National Research Council (CNR) Via G. Celoria 26 I-20133 Milan Italy Institute of Agricultural Biology and Biotechnology National Research Council (CNR) via Bassini 15 I-20133 Milan Italy

Dear Editor, The multiple targeting proteins that have been described in yeast, mammals, and plants have generated intriguing areas of inquiry, including the evolutionary relevance of common proteins shared by different organelles, the mechanisms that determine targeting to each organelle, and, finally, the extent of this phenomenon. Several dual-targeted proteins have been identified in eukaryotic cells.

关键词： MITOCHONDRIA PLANTS CELLS

Rectal hepatoid carcinoma with liver metastases in a patient affected by ulcerative colitis

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Hepatobiliary & Pancreatic Diseases International 2008年第5期7卷 539-543页

作者： Giacomo Borgonovo Francesco Razzetta michela Assalino Emanuela Varaldo Maria Puglisi Paola Ceppa Dipartimento di Discipline Chirurgiche Morfo- logiche e Metodologie Integrate(DiCMI)Universitàdegli Studi di Genova.Largo Benzi8.16132 GenovaItaly

BACKGROUND:Hepatoid tumors(HTs)are rare extra-hepatic neoplasms with the histological features, biochemical profile and,sometimes,even clinical course of hepatocellular *** present a case of rectal hepatoid adenocarcinoma with metachronous liver metastases. METHODS:Four months after total procto-colectomy for a rectal adenocarcinoma(Astler-Coller C2),a 42-year-old man with ulcerative colitis showed hypoechoic masses in the hepatic parenchyma by abdominal ultrasonography. Carcinoembryonic antigen was normal,but alpha- fetoprotein was 32 000μg/***-needle biopsy revealed that liver masses were positive for hepatocellular carcinoma. The patient underwent left hepatectomy and alcoholisation of a small deep nodule in segment 8. RESULTS:Immunohistochemistry and albumin mRNA in situ hybridization suggested that the nodules were metastases of a *** patient was well during the first 6 months and refused any adjuvant *** died from liver failure 19 months after initial diagnosis. CONCLUSIONS:HT is a rare colon *** preoperative diagnosis of this tumor requires a high degree of suspicion,the availability of a panel of immunohistochemical markers,and a certain amount of *** prognosis is poor despite an aggressive andmultimodal therapeutic *** far,none of the hypotheses proposed about the origin and the biology of these tumors is convincing.

关键词： colon cancer ulcerative colitis hepatocellular carcinoma hepatectomy alpha-fetoprotein

Mechanisms involved in selecting and maintaining neuroblastoma cancer stem cell populations, and perspectives for therapeutic targeting

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World Journal of Stem Cells 2021年第7期13卷 685-736页

作者： Antonietta Rosella Farina Lucia Annamaria Cappabianca Veronica Zelli michela Sebastiano Andrew Reay Mackay Department of Applied Clinical and Biotechnological Sciences University of L'AquilaL'Aquila 67100AQItaly

Pediatric neuroblastomas(NBs)are heterogeneous,aggressive,therapy-resistant embryonal tumours that originate from cells of neural crest(NC)origin and in particular neuroblasts committed to the sympathoadrenal progenitor cell *** resistance,post-therapeutic relapse and subsequent metastatic NB progression are driven primarily by cancer stem cell(CSC)-like subpopulations,which through their self-renewing capacity,intermittent and slow cell cycles,drug-resistant and reversibly adaptive plastic phenotypes,represent the most important obstacle to improving therapeutic outcomes in unfavourable *** this review,dedicated to NB CSCs and the prospects for their therapeutic eradication,we initiate with brief descriptions of the unique transient vertebrate embryonic NC structure and salient molecular protagonists involved NC induction,specification,epithelial to mesenchymal transition and migratory behaviour,in order to familiarise the reader with the embryonic cellular and molecular origins and background to *** follow this by introducing NB and the potential NC-derived stem/progenitor cell origins of NBs,before providing a comprehensive review of the salient molecules,signalling pathways,mechanisms,tumour microenvironmental and therapeutic conditions involved in promoting,selecting and maintaining NB CSC subpopulations,and that underpin their therapy-resistant,self-renewing metastatic ***,we review potential therapeutic strategies and future prospects for targeting and eradication of these bastions of NB therapeutic resistance,post-therapeutic relapse and metastatic progression.

关键词： Neural crest Neuroblastoma Cancer stem cells Polyploid giant cancer cells molecular mechanisms Therapeutic targeting Tumour microenvironment