Alessandra Favole | Istituto Zooprofilattico Sperimentale del Piemonte, Liguria e Valle d'Aosta (original) (raw)

Papers by Alessandra Favole

BMC Research Notes, 2021

Objective The spread of bovine spongiform encephalopathy (BSE) agent to small ruminants is still ... more Objective The spread of bovine spongiform encephalopathy (BSE) agent to small ruminants is still a major issue in the surveillance of transmissible spongiform encephalopathies (TSEs). L-type bovine spongiform encephalopathy (L-BSE) is an atypical form of BSE with an unknown zoonotic potential that is transmissible to cattle and small ruminants. Our current knowledge of bovine atypical prion strains in sheep and goat relies only on experimental transmission studies by intracranial inoculation. To assess oral susceptibility of goats to L-BSE, we orally inoculated five goats with cattle L-BSE brain homogenates and investigated pathogenic prion protein (PrP sc ) distribution by an ultrasensitive in vitro conversion assay known as Real-Time Quaking Induced Conversion (RT-QuIC). Results Despite a prolonged observation period of 80 months, all these animals and the uninfected controls did not develop clinical signs referable to TSEs and tested negative by standard diagnostics. Otherwise, R...

Advances in experimental medicine and biology, 2020

Proteoglycans are macromolecules that are essential for the development of cells, human diseases ... more Proteoglycans are macromolecules that are essential for the development of cells, human diseases and malignancies. In particular, chondroitin sulphate proteoglycans (CSPGs) accumulate in tumour stroma and play a key role in tumour growth and invasion by driving multiple oncogenic pathways in tumour cells and promoting crucial interactions in the tumour microenvironment (TME). These pathways involve receptor tyrosine kinase (RTK) signalling via the mitogen-activated protein kinase (MAPK) cascade and integrin signalling via the activation of focal adhesion kinase (FAK), which sustains the activation of extracellular signal-regulated kinases 1/2 (ERK1/2).Human CSPG4 is a type I transmembrane protein that is associated with the growth and progression of human brain tumours. It regulates cell signalling and migration by interacting with components of the extracellular matrix, extracellular ligands, growth factor receptors, intracellular enzymes and structural proteins. Its overexpression...

Aerotecnica Missili & Spazio

“Amyloid Aggregation” is an Italian Space Agency (ASI)-granted project designed to investigate if... more “Amyloid Aggregation” is an Italian Space Agency (ASI)-granted project designed to investigate if and how beta amyloid peptides aggregation is affected by microgravity, in the light of a possible professional risk in astronauts on long-lasting space missions. Researchers of the Istituto Zooprofilattico Sperimentale del Piemonte Liguria e Valle d'Aosta (IZSPLV) and the Istituto Superiore di Sanità (Ist.Sup.San) conceived the project while the Aerospace Logistics Technology Engineering Company (ALTEC) developed the payload. ARGOTEC/Telespazio (UTISS Team) supported safety evaluation, payload manifesting and qualification processes for a safe and efficient delivery, utilization, and integration on board the International Space Station (ISS), and finally recovery of the experimental hardware once returned to Earth. The hardware consisted of 48 special jars containing 6 different time period experimental groups and 2 control groups. In August 2019, during the “BEYOND” mission on board the ISS, astronaut Luca Parmitano activated individual jars according to a defined protocol. At the end of each Incubation Time Periods (ITPs), the samples were transferred in the cold stowage system to stop the aggregation reaction until the analysis after re-entry. Forty-eight identical samples have been prepared and activated in November 2019 on Earth. ALTEC developed jars and packaging for space transportation, qualified and delivered the flight hardware, according to requirements. We here provide an update on the optimization of analytical techniques described in the project specifically intended to warrant an up-to-date, robust and reliable examination of samples that we plan to accomplish in the next months within the proposed timeframe of the project.

PLOS ONE

Scrapie is a fatal neurodegenerative disease of sheep and goats belonging to the group of Transmi... more Scrapie is a fatal neurodegenerative disease of sheep and goats belonging to the group of Transmissible Spongiform Encephalopathy or prion diseases. The EU has adopted mandatory measures for scrapie surveillance to safeguard public and animal health because it is highly contagious and might decimate all genetic susceptible animals in affected flocks. Definite diagnosis of scrapie relies on the detection of the pathological prion protein in brain tissues and there are still no blood biomarkers available for making diagnosis in living animals that can be used for the screening of sheep in scrapie-affected flocks. Neurofilament light (NfL) protein, a valid biomarker for neuronal and axonal damages, can now be easily measured in blood by the ultra-sensitive single molecule array (Simoa) technology. Recent work reported that serum NfL is increased in neurodegenerative diseases, including human prion diseases, but no data are available for scrapie or other animal prion diseases. Here, we found that the median serum NfL concentration in scrapie animals (56.2, IQR 42.2-84.8, n = 9) was more than 15 times higher (p = 0.00084) than that found in control samples (3.4, IQR 3.0-26.3, n = 11). Moreover, serum NfL concentration in scrapie sheep with clinical signs (n = 2; 75.3, 15.7 pg/ml) did not significantly (p = 0.541; t-test) differ from scrapie animals without clinical signs (n = 7; 61.0, 10.7 pg/ml). The receiver operating characteristic (ROC) curve analysis estimated the cutoff value of 31 pg/ml serum NfL for distinguishing scrapie-infected sheep from controls. The application of this cutoff value gives an accuracy of the test of 95% (percent error of 5.23%). These data indicate that the Simoa test for serum NfL might be a useful screening method for detecting preclinical scrapie in living sheep. Finally, the preliminary data reported here need confirmation in large and more structured studies.

Blood

Introduction Multiple myeloma (MM) remains an incurable malignancy despite important recent advan... more Introduction Multiple myeloma (MM) remains an incurable malignancy despite important recent advances in treatments. Neo-vascularization entails a crucial aspect of interactions between neoplastic plasma cells (PCs) and their microenvironment. Without it, MM would be unable to grow and progress, and would probably regress to a low-mass steady-state comparable to monoclonal gammopathy of undetermined significance (MGUS). To overcome drug resistance and improve clinical response to novel therapeutic approaches halting both PC growth and the increased bone marrow (BM) microvascular density are needed. In this setting, monoclonal antibodies against MM-specific cell surface antigens represent a promising therapeutic approach, which is however hampered by a lack of appropriate membrane target structures expressed across all MM cells. The Eph receptors, a large family of receptor tyrosine kinases, have been implicated in many processes involved in malignancy, including alteration of the tum...

Prion - An Overview, 2017

Classical bovine spongiform encephalopathy (C-BSE) is a fatal neurodegenerative disease of cattle... more Classical bovine spongiform encephalopathy (C-BSE) is a fatal neurodegenerative disease of cattle, detected in the United Kingdom and many other countries since the 1980s. The origin of C-BSE is uncertain, but epidemiological studies suggest that the source of this disease was cattle feed prepared from prion-infected animal tissues. To date, cattle populations have been monitored through passive and active surveillance programs. From 2004, two different forms of BSE termed as L-BSE, also known as bovine amyloidotic spongiform encephalopathy (BASE), and H-BSE have been discovered in Italy and France. All these atypical cases have been detected in animals over 8 years of age. To date, there is no comprehensive information about the origin of the atypical BSEs (sporadic vs. acquired). Moreover, there are only very limited data available, concerning the pathogenesis of both atypical forms, as compared to C-BSE. This chapter provides a well-organized overview of what is known about classical and atypical BSE. It will review information on the main epidemiological features, pathogenesis, and the criteria for the routine diagnosis based on rapid tests, histological, immunohistochemical, and Western blot examinations.

Ash Annual Meeting Abstracts, Nov 16, 2012

Abstract 2926 Introduction Angiogenesis plays a central role in the progression of both solid and... more Abstract 2926 Introduction Angiogenesis plays a central role in the progression of both solid and hematological tumors. In particular, in multiple myeloma (MM) the critical role of bone marrow (BM) microenvironment and angiogenesis has been well documented. The past decade has witnessed a dramatic improvement in the therapeutic options in MM. However, the disease remains incurable, underscoring the need for continued efforts towards understanding MM biology and exploitation of novel therapeutic approaches. In this setting, monoclonal antibodies against myeloma-specific cell surface antigens represent a promising therapeutic approach, which is however hampered by a lack of appropriate target structures expressed across all pathogenic myeloma cells. The Eph receptors, a large family of receptor tyrosine kinases (RTKs) activated by ephrins binding, have been implicated in many processes involved in malignancy, including alteration of the tumor microenvironment and in angiogenesis, in both of which EpHA3 likely plays an active role. Aberrant expression of EpHA3 is seen in many types of hematolologic malignancies (some leukemic cell lines, T-cell lymphoma, acute lymphoblastic leukemia, myeloproliferative neoplasms) although it is not expressed ubiquitously. Finally, the over-expression of Eph is believed to be sufficient to confer tumorigenic potential although probably further mechanisms can occur to abnormally activate the receptor. Basing on the role of EpHA3 in haematological malignancies, a first-in-class engineered IgG1 antibody targeting the EpHA (KB004) was developed and it is now under phase I clinical trials in USA and Australia for the treatment of EpHA3 overexpressing hematological myeloid malignancies refractory to conventional treatment. We investigated the EpHA3 role and its preferential membrane–bound by GPI linker ligand EFNA5, in MM patients in order to define EpHA3 as new molecular target for a novel therapeutic approach with a specific anti EpHA3 monoclonal antibody. The EpHA3 expression has been studied through a comparative proteomic analysis between BM endothelial cells (ECs) of patients with MM (MMECs) or with monoclonal gammopathy of undetermined significance (MGECs), of control subjects (normal ECs) and in MM cell lines. Methods After written informed consent, BM aspirates have been collected from 20 MM and 4 MGUS patients. Normal ECs were derived from 3 BM aspirates of subjects with anemia due to iron or vitamin B12 deficiency. We analyzed the expression levels of EpHA3 in normal ECs, MGECs and MMECs and MM cell lines evaluating the mRNA and protein levels by RT-qPCR and by WB coupled to ImmunoFluorescence analysis. The biological effects of EpHA3 targeting in MMECs have been studied silencing the EpHA3 mRNA in MMECs and testing them at 72h after silencing in series of functinal assays including viability assay by trypan blue exclusion staining and by in vitro angiogenesis assay followed by measurement of mesh areas and vessel length. Moreover, we studied EFNA5 mRNA expression levels in Normal ECs, MGECs and MMECs and in MM cell lines by PCR. Results Our data showed that EpHA3 mRNA levels are progressively increased from ECs to MGECs reaching the highest values in MMECs. Subsequent analysis by WB and immunofluorescence confirmed EpHA3 protein upregulation among the different EC types. The MMECs in which EpHA3 has been silenced revealed a protein level reduction of approximately 60% when compared to the control. We could not detect major viability defects. Furthermore, in vitro angiogenesis inhibition was marginal when compared to the not silenced counterpart. To know whether EpHA3 may impact not only MM angiogenesis but also plasma cells, three MM cell lines were studied for the EpHA3 expression. We found the plasma cell lines gave constant over expression of EpHA3. Finally, the preliminary data regarding EFNA5 mRNA expression level showed it is expressed in either MMECs and MM plasma cell lines. The evaluation of KB004 effect on MMECs in term of apoptosis induction and in vitro tube formation inhibition, as well as the analysis of EpHA3 levels in primary MM plasma cells are in progress. Conclusions From this study we expect to characterize the role of the EpHA3in MM patients and to provide experimental evidences supporting the possibility of using EpHA3 as a new molecular target for MM by proving the in vitro efficacy of a monoclonal antibody to target the angiogenesis of MM. Disclosures: No relevant conflicts of interest to declare.

Ash Annual Meeting Abstracts, Nov 16, 2012

Abstract 1761 Background: Mutation(s) of the JAK2 gene (V617F) has been described in a significan... more Abstract 1761 Background: Mutation(s) of the JAK2 gene (V617F) has been described in a significant proportion of Philadelphia negative MPN patients and its detection is now a cornerstone in the diagnostic algorithm. The method most frequently used for measuring the distribution of cell populations is based on JAK2 sequencing and Q-PCR. Therefore the chance of distinguishing the JAK2 wild type or mutated population at the single-cell level still represents a challenge. The aim of the study was to developed a novel assay based on peptide nucleic acid (PNA) technology coupled to immuno-fluorescence microscopy (PNA-FISH) for the specific detection at a single cell level of JAK2-mutation thus improving both the diagnostic resolution and the study of clonal prevalence. Methods: We designed a fluorescently-labelled PNA probe, coupled to FISH technology, which allows to distinguish with a high degree of specificity between CD34+ progenitor stem cells harbouring the mutated (V617F) or the wild type form of JAK2. CD34+ cells were enriched from 24 PV patients (5 of them were selected for the absence of JAK2V617F), 13 PMF (10 with the mutation and 3 JAK2 wild type) and 6 ET patients (2 of them were wild type). In addition 20 BM samples were collected from healthy donors and used as control. Patients were a priori found to be either positive or negative for the JAK2V617F mutation by standard sequencing and by Q-PCR. CD34+ progenitors cells were enriched by MACS and then cytospun onto slides and hybridized with human species-specific fluorescinated 15 base pairs (bp)-long oligo-PNA, specifically recognizing the human JAK2 sequence surrounding the nucleotide at position 1849, which is responsible for the V617F substitution (JAK2V617F/PNA). Slides were analyzed by fluorescence confocal microscopy. Results: The analysis revealed that among JAK2V617F PV patients the distribution pattern is fairly similar to that reported by Scott and colleagues in 2006 analyzing JAK2V617F in colonies. We found, with a rather wide variability occurring among patients, a percentage of mutated CD34+ cells ranging from 40% to 100% in PV patients, from 15% to 80% in ET and from 25% to 100% in PMF. These findings are in agreement with previous data reporting that a variable proportion of progenitors from patients affected by JAK2V617F positive PV are capable of generating JAK2V617F negative colonies. In addition these data indicate that fluorescinated JAK2V617F/PNA probe displays a very high specificity towards a single base-pair mismatch. Interestingly, when evaluating the presence of JAK2V617Fpositive cells collected from 3 JAK2 wild type subjects defined by sequencing and by Q-PCR, we identified a small percentage of cells positive for the JAK2V617F/PNA staining. However, this apply only to patients with PV but not to PMF and ET patients. Quantitatively, this percentage did not exceed 3% of the CD34+ cell population indicating a high level of sensitivity of the procedure since the PNA/FISH technique may detect JAK2V617F-positive cells within a CD34+ population isolated from patients reported as JAK2V617Fnegative by standard approaches. Importantly, the lack of positivity detected in CD34 positive cells from 20 healthy subjects demonstrates a high specificity of this method. Conclusions: JAK2V617F/PNA-FISH method displays high specificity and reliability in discriminating cell subpopulations harbouring the JAK2V617F mutation. In addition, it allows to analyze the CD34+ population at the single cell level, avoiding the time consuming analysis of hematopoietic colonies. In addition, this approach allows to monitor longitudinally the evolution of a defined cell population over time in MPNs and the characterization of the CD34+ compartment in patients with MPNs which still represents a challenging issue. Disclosures: No relevant conflicts of interest to declare.

Ash Annual Meeting Abstracts, Nov 16, 2012

Abstract 4616 Background: The Sprouty family proteins, including Spred, were originally identifie... more Abstract 4616 Background: The Sprouty family proteins, including Spred, were originally identified in Drosophila melanogaster as an antagonist of Breathless, the ortholog of Fibroblast Growth Factor Receptor (FGFR) in mammals. These proteins are inducible inhibitors of signalling induced by receptor tyrosine kinases. Their main function is to deregulate the RAS/MAPK and RAS/RAF/ERK signal pathways by physically interacting. The role of Spread proteins in haematological malignancies is still not been clarified. Aim: The aim of this study was to investigate a possible involvement of Spred in maintaining the aberrant TK signalling in patients affected by acute myeloid leukaemia (AML). Methods: Bone marrow (BM) cells were collected from 82 AML patients at diagnosis and 10 BM samples from healthy donors as control. In addition 20 patients were analyzed during follow-up at the time haematological remission. All the patients included had been characterized at the cytogenetic level by conventional karyotyping, and screened by reverse transcriptase-PCR for the presence of the most frequent fusion transcripts or mutations. We analysed Spread mRNA expression by RQ-PCR and the protein by Western blot and immunofluorescence assay. In addition gain of function experiments were performed by transfecting with Spred coding sequence different leukemic cells lines lacking Spred activity. Results: We found that Spread mRNA levels were significantly decreased in AML compared to healthy subjects (2−DDct = 0,1 in AML compared to 0,6 in controls) (p&amp;amp;amp;amp;amp;amp;lt;0,001). Western blot and immunofluorescence confirmed the Spred downregulation at the protein level. Spread expression did not correlate to FAB subtypes or cytogenetic risk. Interestingly, when patients obtained the complete hematological remission (HR) the levels of Spred mRNA and protein were significantly upregulated to a level similar to healthy subjects (p=0.1 between HR and normal controls). Transfection experiments of Kasumi cell line with Spred coding sequence resulted in a decrease of proliferation of 53%, a colony growth reduction of 50% and a significant increase of apoptosis (p&amp;amp;amp;amp;amp;amp;lt;0.001). Finally, in order to establish the molecular basis for Spred1 deregulation we analyzed the expression levels of miRNA-21 which is a known regulator of Spred and we found a correlation between the expression of miRNA-21 and Spred. Conclusions: A decreased expression and activity of Spred, a negative regulator of TK activity through Ras pathway, is implicated in sustaining the oncogenetic signalling and abnormal proliferation induced by tyrosine kinase proteins in acute myeloid leukemia patients. Disclosures: No relevant conflicts of interest to declare.

Hematology, 2005

The combined use of bone marrow histopathology, biomarkers and clinical features has the potentia... more The combined use of bone marrow histopathology, biomarkers and clinical features has the potential to diagnose, stage and distinguish early and overt stages of ET, PV and idiopathic myelofibrosis, that has an important impact on prognosis and treatment of MPD patients. As the extension of the PVSG and WHO for ET, PV and agnogenic myeloid metaplasia (AMM), a new set of European clinical and pathological (ECP) criteria clearly distinct true ET from early or latent PV mimicking true ET, overt and advanced polycythemia vera (PV), and from thrombocythemia associated with prefibotic, early fibrotic stages of chronic megakaryocytic granulocytic metaplasia (CMGM) or chronic idiopathic myelofibrosis (CIMF). Cases of atypical MPD and masked PV are usually overlooked by clinicians and pathologists. Bone marrow biopsy will not differentiate between post-PV myelofibrosis versus so-called classical agnogenic myeloid metaplasia. The recent discovery of the JAK2 V617F mutation can readily explain the trilinear megakaryocytic, erythroid and granulocytic proliferation in the bone marrow, but also the etiology of the platelet-mediated microvascular thrombotic complications at increased platelet counts and red cell mass in essential thrombocythemia and polycythemia vera.

Scientific Reports

Since 2005, two cases of natural bovine spongiform encephalopathies (BSE) have been reported in g... more Since 2005, two cases of natural bovine spongiform encephalopathies (BSE) have been reported in goats. Furthermore, experimental transmissions of classical (C-BSE) and atypical (L-BSE) forms of BSE in goats were also reported. To minimize further spreading of prion diseases in small ruminants the development of a highly sensitive and specific test for ante-mortem detection of infected animals would be of great value. Recent studies reported high diagnostic value of a second generation of cerebrospinal fluid (CSF) Real-Time Quaking-Induced Conversion (RT-QuIC) assay across a wide spectrum of human prions. Here, we applied this improved QuIC (IQ-CSF) for highly efficient detection of TSEs prion protein in goat cerebrospinal fluid. IQ-CSF sensitivity and specificity were evaluated on CSF samples collected at disease endpoint from goats naturally and experimentally infected with scrapie or bovine isolates of C-Bse and L-Bse, respectively. Next, CsF samples collected from L-Bse infected goats during pre-symptomatic stage were also analysed. PrP L-Bse associated seeding activity was detected at early time points after experimental inoculation, with an average time of 439 days before clinical symptoms appeared. taken together these data are indicative of the great potential of this in vitro prion amplification assay as ante-mortem tse test for live and asymptomatic small ruminants. Transmissible Spongiform Encephalopathies (TSEs) or prion diseases are fatal neurodegenerative diseases that include Creutzfeldt-Jakob disease (CJD) in humans, bovine spongiform encephalopathy (BSE) in cattle, scrapie in sheep and chronic wasting disease (CWD) in cervids. The infectious agent responsible for these diseases, the prion, appears to be composed primarily of an abnormal, misfolded and oligomeric (PrP Sc) form of the cellular prion protein (PrP C) 1,2. PrP Sc induces the polymerization and conformational conversion of PrP C to its infectious form 3-5 or, in a variety of in vitro conversion assays 6 , into alternative forms of the prion protein which are, similarly to PrP Sc , partially resistant to digestion with proteases (PrP Res). The pathogenic isoform of the prion protein is therefore a marker associated to TSEs which acts as a seed allowing the ultrasensitive detection of PrP Sc using in vitro prion amplification reactions such as Real-Time Quaking Induced Conversion (RT-QuIC) 7. The spread of BSE agent to small ruminants is a major issue in the surveillance of TSEs because BSE passage into a new host may change strain properties, make it difficult to recognize the original strain, and increasing the risk of epidemic spread 8. Since 2005, two natural BSE cases have been reported in goats 9,10. Furthermore,

Journal of clinical microbiology, Jan 21, 2015

Statutory surveillance of bovine spongiform encephalopathy (BSE) indicates that cattle are suscep... more Statutory surveillance of bovine spongiform encephalopathy (BSE) indicates that cattle are susceptible to both classical (C-BSE) and atypical forms of BSE. Atypical forms of BSE appear to be sporadic and thus may never be eradicated. A major challenge for prion surveillance is the lack of sufficiently practical and sensitive tests for routine BSE detection and strain discrimination. The RT-QuIC test, which is based on prion-seeded fibrillization of recombinant prion protein (rPrP(Sen)), is known to be highly specific and sensitive for detection of multiple human and animal prion diseases, but not BSE. Here, we tested brain tissue from C-BSE- and atypical L-type bovine spongiform encephalopathy (L-type BSE or L-BSE) affected cattle by RT-QuIC and found that both BSE forms can be detected and distinguished using particular rPrP(Sen) substrates. Specifically, L-BSE was detected using multiple rPrP(Sen) substrates while C-BSE was much more selective. This substrate-based approach sugges...

Translational oncology, 2012

Defects in HLA class I antigen-processing machinery (APM) component expression and/or function ar... more Defects in HLA class I antigen-processing machinery (APM) component expression and/or function are frequent in human tumors. These defects may provide tumor cells with a mechanism to escape from recognition and destruction by HLA class I antigen-restricted, tumor antigen-specific cytotoxic T cells. However, expression and functional properties of MHC class I antigens and APM components in malignant cells in other animal species have been investigated to a limited extent. However, this information can contribute to our understanding of the mechanisms underlying the association of MHC class I antigen and APM component defects with malignant transformation of cells and to identify animal models to validate targeted therapies to correct these defects. To overcome this limitation in the present study, we have investigated the expression of the catalytic subunits of proteasome (Y, X, and Z) and of immunoproteasome (LMP2, LMP7, and LMP10) as well as of MHC class I heavy chain (HC) in 25 pr...

Veterinary Research Communications, 2007

Accornero, P., Luvar�, S., Favole, A., Macchi, E., Motta, M., Baratta, M., 2007. Biological role ... more Accornero, P., Luvar�, S., Favole, A., Macchi, E., Motta, M., Baratta, M., 2007. Biological role of the HGF/MET ligand/receptor couple in bovine mammary epithelial cells. Veterinary Research Communications, 31(Suppl. 1), 161–164

Leukemia Research, 2013

Chronic myelomonocytic leukemia (CMML) is a clonal disorder sharing features of myelodysplastic s... more Chronic myelomonocytic leukemia (CMML) is a clonal disorder sharing features of myelodysplastic syndromes and chronic myeloproliferative neoplasms. Although rare chromosomal aberrations and point mutations are reported in CMML, the molecular defects underlying CMML are largely unknown. ROS1 encodes a tyrosine kinase that is abnormally expressed and translocated in brain and lung cancers. In this study we show that ROS1 is abnormally activated in the CD34+ compartment of approximately 70% of CMML patients resulting in the activation of the Erk/Akt pathways through the Grb2/SOS complex thus revealing a central oncogenic role for ROS1 in CMML which might represent a molecular target.

European Journal of Cancer, 2012

Cancer Research, 2011

Cell surface chondroitin sulfate proteoglycan 4 (CSPG4) is an attractive target for antibody-base... more Cell surface chondroitin sulfate proteoglycan 4 (CSPG4) is an attractive target for antibody-based cancer immunotherapy because of its role in tumor cell biology, its high expression on malignant cells including cancer-initiating cells, and its restricted distribution in normal tissues. The clinical use of CSPG4 has been hampered by the lack of a CSPG4-specific chimeric, humanized, or fully human monoclonal antibody. To overcome this limitation, we generated a CSPG4-specific fully human single-chain antibody termed scFv-FcC21 and characterized its specificity and antitumor activity. Viable CSPG4(+) melanoma cells were used in a screen of a human scFv phage display library that included CDR3 engineered to optimize antibody binding sites. The scFv antibody isolated was then recombinantly engineered with a human immunoglobulin G1 Fc region to construct the fully human antibody scFv-FcC21, which recognized tumors of neuroectodermal origin, various types of carcinomas, mesotheliomas, and sarcomas as well as myeloid leukemias. scFv-FcC21 inhibited in vitro growth and migration of tumor cells and in vivo growth of human tumor xenografts. These effects were mediated by inhibition of the activation of extracellular signal-regulated kinase and focal adhesion kinase signaling pathways that are critical for tumor cell growth and migration, respectively. Our findings define the CSPG4-specific fully human scFv-FcC21 antibody as a candidate therapeutic agent to target the many types of tumors that express CSPG4.

BMC Research Notes, 2021

Advances in experimental medicine and biology, 2020

Aerotecnica Missili & Spazio

PLOS ONE

Blood

Prion - An Overview, 2017

Ash Annual Meeting Abstracts, Nov 16, 2012

Hematology, 2005

Scientific Reports

Journal of clinical microbiology, Jan 21, 2015

Translational oncology, 2012

Veterinary Research Communications, 2007

Leukemia Research, 2013

European Journal of Cancer, 2012

Cancer Research, 2011