A5042197595- Virus-based gene therapy research
- CRISPR and Genetic Engineering
- CAR-T cell therapy research
- RNA Interference and Gene Delivery
- Viral Infectious Diseases and Gene Expression in Insects
- T-cell and B-cell Immunology
- Genetic and phenotypic traits in livestock
- Genetic Mapping and Diversity in Plants and Animals
- Immune Cell Function and Interaction
- HIV Research and Treatment
- RNA Research and Splicing
- Neuroblastoma Research and Treatments
- Gene expression and cancer classification
- Molecular Biology Techniques and Applications
- Viral Infections and Immunology Research
- Bioinformatics and Genomic Networks
- Viral gastroenteritis research and epidemiology
- Histone Deacetylase Inhibitors Research
- Protein Degradation and Inhibitors
- RNA and protein synthesis mechanisms
- Genetic factors in colorectal cancer
- Immunotherapy and Immune Responses
- Genomics and Phylogenetic Studies
- Monoclonal and Polyclonal Antibodies Research
- Bacteriophages and microbial interactions
Protagen (Germany)
2023-2024
Gene Bridges (Germany)
2017-2022
German Cancer Research Center
2012-2021
Heidelberg University
2012-2021
National Center for Tumor Diseases
2012-2021
University of Bologna
2005-2012
Polytechnic University of Bari
2009
Policlinico S.Orsola-Malpighi
2005
Wiskott-Aldrich syndrome gene therapy is feasible, but γ-retroviral vectors contribute a substantial risk of leukemogenesis.
Recombinant adeno-associated viral vectors (rAAV) currently constitute a real therapeutic strategy for the sustained correction of diverse genetic conditions. Though wealth preclinical and clinical studies have been conducted with rAAV, oncogenic potential these is still controversial, particularly when considering liver-directed gene therapy. Few recent discovery incomplete wild-type AAV2 genomes integrated in human hepatocellular carcinoma biopsies raised concerns on rAAV safety. In...
Abstract Unbiased dissection of T-cell receptor (TCR) repertoire diversity at the nucleotide level could provide important insights into human immunity. Here we show that TCR ligation-anchored-magnetically captured PCR (TCR-LA-MC PCR) identifies α- and β-chain without sequence-associated or quantitative restrictions in healthy diseased conditions. TCR-LA-MC convergent recombination events, classifies different stages cutaneous lymphoma vivo demonstrates reactivation after vitro...
A previously published clinical trial demonstrated the benefit of autologous CD34+ cells transduced with a selfinactivating lentiviral vector (HPV569) containing an engineered β-globin gene (βA-T87Q-globin) in subject β thalassemia major. This has been modified to increase transduction efficacy without compromising safety. In vitro analyses indicated that changes resulted both increased titers (3 4 fold) and (2 3 fold). An vivo study which 58 β-thalassemic mice were transplanted vector- or...
Antigen exposure and differentiation phenotype influence long-term persistence of memory T cells after hematopoietic stem cell transplant.
HIV-1 recurrently targets active genes and integrates in the proximity of nuclear pore compartment CD4+ T cells. However, genomic features these relevance their transcriptional activity for integration have so far remained unclear. Here we show that targeted are proximal to super-enhancer elements they cluster specific spatial compartments cell nucleus. We further gene clusters acquire location during activation The clustering along with major determinants Our results provide evidence...
Tumour onset and progression are due to the accumulation of genomic lesions, which alter gene expression ultimately proteome activities. These lesions thought affect primarily transcriptional control expression. In present study, we aimed at evaluating genome-wide occurrence alterations in translational exploiting an isogenic, phenotypically validated cellular model colorectal cancer (CRC) transition from invasive carcinoma metastasis. this model, microarray profiling shows that changes...
Combining different approaches (resequencing of portions 54 obesity candidate genes, literature mining for pig markers associated with fat deposition or related traits in 77 and silico porcine expressed sequence tags other sequences available databases), we identified analyzed 736 SNP within genes to identify back thickness (BFT) Italian Large White sows. Animals were chosen using a selective genotyping approach according their EBV BFT (276 most negative 279 positive EBV) population ≈ 12,000...
Abstract Regulatory CD4 + T cells (Treg) prevent tumor clearance by conventional (Tconv) comprising a major obstacle of cancer immune-surveillance. Hitherto, the mechanisms Treg repertoire formation in human cancers remain largely unclear. Here, we analyze clonal origin breast patients using T-Cell Receptor and single-cell transcriptome sequencing. While peripheral blood tumors are clonally distinct, Tconv clones, including tumor-antigen reactive effectors (Teff), detected both compartments....
In vivo reprogramming of somatic cells into induced pluripotent stem (iPSC) holds vast potential for basic research and regenerative medicine. However, it remains hampered by a need vectors to express factors (Oct-3/4, Klf4, Sox2, c-Myc; OKSM) in selected organs. Here, we report OKSM delivery based on pseudotyped Adeno-associated virus (AAV). Using the AAV-DJ capsid, could robustly reprogram mouse embryonic fibroblasts with low vector doses. Swapping AAV8 permitted efficiently adult mice...
We developed an anti-gene peptide nucleic acid (PNA) for selective inhibition of MYCN transcription in neuroblastoma cells, targeted against a unique sequence the antisense DNA strand exon 2 and linked at its NH(2) terminus to nuclear localization signal peptide. Fluorescence microscopy showed specific delivery PNA six human cell lines: GI-LI-N IMR-32 (MYCN-amplified/overexpressed); SJ-N-KP NB-100 (MYCN-unamplified/low-expressed); GI-CA-N GI-ME-N (MYCN-unamplified/unexpressed)....
A hierarchically organized cell compartment drives colorectal cancer (CRC) progression. Genetic barcoding allows monitoring of the clonal output tumorigenic cells without prospective isolation. In this study, we asked whether tumor clone-initiating (TcICs) were genetically heterogeneous and differences in self-renewal activation reflected differential kinetics among individual subclones or functional hierarchies within subclones. Monitoring genomic subclone three patient tumors corresponding...
Integration site profiling and clonality analysis of viral vector distribution in gene therapy is a key factor to monitor the fate gene-corrected cells, assess risk malignant transformation, establish biosafety. We developed Genome Site Analysis Pipeline (GENE-IS) for highly time-efficient accurate detection next-generation sequencing (NGS)-based integration sites (ISs) data. It first available tool with dual mode that allows IS both data generated by PCR-based methods, such as linear...
We developed an antisense peptide nucleic acid (PNA) targeted against a unique sequence in the terminus of 5'-UTR N-myc, designed for selective inhibition NMYC neuroblastoma cells. Fluorescent microscopy showed carrier-free delivery PNA to two human neuro-blastoma cell lines: GI-LI-N (N-myc-amplified) and GI-CA-N (N-myc-unamplified). Only former, treatment determined 70% cell-viability reduction (at 48 h). In N-myc-amplified cells, NMYC-translation (Western blotting), accumulation cells G1,...
Abstract Lentivirus vectors are effective for treatment of genetic disease and cancer, however, vector related insertional mutagenesis genotoxicity is concern currently available safety models not reliably predictive in humans. We have developed h InGeTox as the first human vitro platform that uses induced pluripotent stem cells their hepatocyte like derivatives to further understand LV host interaction evaluation design. To characterise genotoxic association, we used LTR SIN configuration...
Chronic granulomatous disease (CGD) is a primary immunodeficiency characterized by impaired antimicrobial activity in phagocytic cells. As monogenic affecting the hematopoietic system, CGD amenable to gene therapy. Indeed phase I/II clinical trial, we demonstrated transient resolution of bacterial and fungal infections. However, therapeutic benefit was compromised occurrence clonal dominance malignant transformation demanding alternative vectors with equal efficacy but safety-improved...
We have previously shown that injury-induced neointima formation was rescued by adenoviral-Nogo-B gene delivery. Integrase-competent lentiviral vectors (ICLV) are efficient at delivery to vascular cells but present a risk of insertional mutagenesis. Conversely, integrase-deficient (IDLV) offer additional benefits through reduced mutagenesis risk, this has not been evaluated in the context transfer. Here, we investigated performance and genetic safety both counterparts primary human smooth...
Reactivation of latent viruses such as human cytomegalovirus (HCMV) after allogeneic hematopoietic stem cell transplantation (HSCT) results in high morbidity and mortality. Effective immunization against HCMV shortly allo-HSCT is an unmet clinical need due to delayed adaptive T development. Donor-derived dendritic cells (DCs) have a critical participation stimulation naïve immune reconstitution, therefore adoptive DC therapy could be used protect patients HSCT. However, previous methods for...
Lentiviral vectors (LV) are attractive for permanent and effective gene therapy. However, integration into the host genome can cause insertional mutagenesis highlighting importance of understanding LV integration. Insertion site (IS) tethering is believed to involve cellular proteins such as PSIP1/LEDGF/p75, which binds virus pre-integration complexes (PICs) helping target genome. Transcription factors (TF) that bind both vector LTR also suspected influential this. To determine role TF in...
High-throughput sequencing technologies have exposed the possibilities for in-depth evaluation of T-cell receptor (TCR) repertoires. These studies are highly relevant to gain insights into human adaptive immunity and decipher composition diversity antigen receptors in physiological disease conditions. The major objective TCR data analysis is identification V, D J gene segments, complementarity-determining region 3 (CDR3) sequence extraction clonality analysis. With advancement technologies,...
High-throughput integration site (IS) analysis of wild-type adeno-associated virus type 2 (wtAAV2) in human dermal fibroblasts (HDFs) and HeLa cells revealed that juxtaposition a Rep binding (RBS) terminal resolution (trs)-like motif leads to 4-fold-increased probability wtAAV integration. Electrophoretic mobility shift assays (EMSAs) confirmed off-target RBSs. For the first time, we show protein nicking activity, highlighting importance substrate for Rep-mediated