Secreted Gal-3BP is a novel promising target for non-internalizing Antibody–Drug Conjugates

Abstract Galectin-3-binding protein (Gal-3BP) has been identified as a cancer and metastasis-associated, secreted protein that is expressed by the large majority of cancers. The present study describes a special type of non-internalizing antibody-drug-conjugates that specifically target Gal-3BP. Here, we show that the humanized 1959 antibody, which specifically recognizes secreted Gal-3BP, selectively localized around tumor but not normal cells. A site specific disulfide linkage with thiol-maytansinoids to unpaired cysteine residues of 1959, resulting in a drug-antibody ratio of 2, yielded an ADC product, which cured A375m melanoma bearing mice. ADC products based on the non-internalizing 1959 antibody may be useful for the treatment of several human malignancies, as the cognate antigen is abundantly expressed and secreted by several cancers, while being present at low levels in most normal adult tissues

Injury-specific factors in the cerebrospinal fluid regulate astrocyte plasticity in the human brain

The glial environment influences neurological disease progression, yet much of our knowledge still relies on preclinical animal studies, especially regarding astrocyte heterogeneity. In murine models of traumatic brain injury, beneficial functions of proliferating reactive astrocytes on disease outcome have been unraveled, but little is known regarding if and when they are present in human brain pathology. Here we examined a broad spectrum of pathologies with and without intracerebral hemorrhage and found a striking correlation between lesions involving blood-brain barrier rupture and astrocyte proliferation that was further corroborated in an assay probing for neural stem cell potential. Most importantly, proteomic analysis unraveled a crucial signaling pathway regulating this astrocyte plasticity with GALECTIN3 as a novel marker for proliferating astrocytes and the GALECTIN3-binding protein LGALS3BP as a functional hub mediating astrocyte proliferation and neurosphere formation. Taken together, this work identifies a therapeutically relevant astrocyte response and their molecular regulators in different pathologies affecting the human cerebral cortex. Intracerebral hemorrhage triggers astrocyte proliferation and mediates neural stem cell potential via Galectin3 signaling

Patterns of K-ras mutation in colorectal carcinomas from Iran and Italy (a Gruppo Oncologico dell'Italia Meridionale study): influence of microsatellite instability status and country of origin.

Background: K-ras mutations are a key step in colorectal cancer progression. Such mutations have been widely studied in case series from Western countries but there are few data on the rate and spectrum of mutations in tumors from countries where the epidemiological features of the disease are different. Patients and methods: Tumor samples from 182 Iranian colorectal cancer patients (170 sporadic cases and 12 HNPCC cases) were screened for K-ras mutations at codons 12, 13 and 61 by sequencing analysis. The cases were also characterized for microsatellite instability at mononucleotide repeats by PCR and fragment analysis, and classified according to microsatellite instability status. The frequency and the spectrum of K-ras mutations were compared with those observed in a series of colorectal cancer patients from Italy. Results: K-ras mutations were observed in 68/182 (37.4%) cases. Mutation frequencies were similar in HNPCC-associated, sporadic MSI-H and sporadic microsatellite-stable (MSS) tumors. However, the G13D substitution was more frequent in HNPCC (3/ 4, 75%) and sporadic MSI- H (7/11, 63.6%) tumors compared to sporadic MSS tumors (11/ 53, 20.4%) (P < 0.01). Comparison of mutations in the two series from Iran and Italy showed a significantly higher frequency of G13D among Italian patients. Conclusions: While the frequency of K-ras mutations could be similar, the mutational spectrum could be differentially influenced by genetic and environmental factors

The LOFAR view of intergalactic magnetic fields with giant radio galaxies

Context. Giant radio galaxies (GRGs) are physically large radio sources that extend well beyond their host galaxy environment. Their polarization properties are affected by the poorly constrained magnetic field that permeates the intergalactic medium on megaparsec scales. A low frequency ( Aims: Here we investigate the polarization properties and Faraday rotation measure (RM) of a catalog of GRGs detected in the LOFAR Two-meter Sky Survey. This is the first low frequency polarization study of a large sample of radio galaxies that were selected on their physical size. We explore the magneto-ionic properties of their under-dense environment and probe intergalactic magnetic fields using the Faraday rotation properties of their radio lobes. LOFAR is a key instrument for this kind of analysis because it can probe small amounts of Faraday dispersion (-2), which are associated with weak magnetic fields and low thermal gas densities. Methods: We used RM synthesis in the 120-168 MHz band to search for polarized emission and to derive the RM and fractional polarization of each detected source component. We study the depolarization between 1.4 GHz and 144 MHz using images from the NRAO VLA Sky Survey. We investigate the correlation of the detection rate, the RM difference between the lobes, and the depolarization with different parameters as follows: the angular and linear size of the sources and the projected distance from the closest foreground galaxy cluster. In our sample, we also included 3C 236, which is one of the largest radio galaxies known. Results: From a sample of 240 GRGs, we detected 37 sources in polarization, all of which have a total flux density above 56 mJy. We detected significant RM differences between the lobes, which would be inaccessible at gigahertz frequencies, with a median value of ∼1 rad m-2. The fractional polarization of the detected GRGs at 1.4 GHz and 144 MHz is consistent with a small amount of Faraday depolarization (a Faraday dispersion -2). Our analysis shows that the lobes are expanding into a low-density (-5 cm-3) local environment that is permeated by weak magnetic fields (500. In general, this work demonstrates the ability of LOFAR to quantify the rarefied environments in which these GRGs exist and highlights them as an excellent statistical sample to use as high precision probes of magnetic fields in the intergalactic medium and the Milky Way

Spectral study of the diffuse synchrotron source in the galaxy cluster Abell 523

The galaxy cluster Abell 523 (A523) hosts an extended diffuse synchrotron source historically classified as a radio halo. Its radio power at 1.4 GHz makes it one of the most significant outliers in the scaling relations between observables derived from multiwavelength observations of galaxy clusters: it has a morphology that is different and offset from the thermal gas, and it has polarized emission at 1.4 GHz typically difficult to observe for this class of sources. A magnetic field fluctuating on large spatial scales (similar to 1 Mpc) can explain these peculiarities but the formation mechanism for this source is not yet completely clear. To investigate its formation mechanism, we present new observations obtained with the LOw Frequency ARray at 120-168 MHz and the Jansky Very Large Array at 1-2 GHz, which allow us to study the spectral index distribution of this source. According to our data the source is observed to be more extended at 144 MHz than previously inferred at 1.4 GHz, with a total size of about 1.8 Mpc and a flux density S-144 MHz = (1.52 +/- 0.31) Jy. The spectral index distribution of the source is patchy with an average spectral index alpha similar to 1.2 between 144 MHz and 1.410 GHz, while an integrated spectral index alpha similar to 2.1 has been obtained between 1.410 and 1.782 GHz. A previously unseen patch of steep spectrum emission is clearly detected at 144 MHz in the south of the cluster. Overall, our findings suggest that we are observing an overlapping of different structures, powered by the turbulence associated with the primary and a possible secondary merger.Peer reviewe

Galectin-3 binding protein stimulated IL-6 expression is impeded by antibody intervention in SARS-CoV-2 susceptible cell lines

COVID‑19 is the global pandemic that affected our population in the past 2 years. Considerable research has been done to better understand the pathophysiology of this disease and to identify new therapeutic targets, especially for severe cases. Galectin‑3 (Gal‑3) is a receptor present at the surface of different cell types, namely epithelial and inflammatory cells, which has been described as a severity marker in COVID‑19. The activation of Gal‑3 through its binding protein (Gal‑3BP) is directly linked to the production of pro‑inflammatory cytokines that contribute for the cytokine storm (CS) observed in severe COVID‑19 patients. Here, we show that D2, a recombinant fragment of the lectin‑binding region of Gal‑3BP was able to stimulate the expression of IL‑6 in colon and lung epithelial cell lines in β‑galactoside dependent manner. We further show that D2‑induced IL‑6 augmentation was reduced by the anti‑Gal‑3BP monoclonal antibody 1959. Our data confirm and extend prior findings of Gal‑3BP mediated IL‑6 induction, enlightening the potential of its antibody‑mediated s blockage for the prevention and treatment of CS and severe disease in COVID‑19 patients

Utilizzo terapeutico di un dominio della proteina umana LGALS3BP denominato D2 nell’arteriopatia obliterante periferica e nelle ulcere cutanee croniche. Therapeutic use of a domain of the human LGALS3BP protein called D2 in peripheral arterial disease and chronic skin ulcers.

La presente invenzione riguarda l’utilizzo farmacologico di uno specifico dominio della proteina umana LGALS3BP (nota anche come Mac-2 BP o Gal-3BP o 90K) caratterizzato dalla sequenza amminoacidica riportata nella tabella 1 e qui denominato D2. Gli inventori hanno scoperto che tale dominio è in grado di mediare processi fisiologici, incluso la rigenerazione tissutale e l’angiogenesi, che sono essenziali per la cura delle patologie ischemiche e la guarigione delle ulcere diabetiche del piede. Per tali patologie non esistono attualmente farmaci efficaci e specifici, compresi gli antiinfiammatori. Nella presente invenzione viene dimostrato che la somministrazione di D2 a dosi inferiori a 1 mg, porta a una significativa riduzione del tempo di guarigione di ferite sperimentali nell’animale e a un aumento del flusso sanguigno in un modello murino di ischemia degli arti posteriori. La sostituzione di alcuni aminoacidi con altri amino acidi (&lt; 5% del totale) nella sequenza di D2 non porta a sostanziali modifiche dell’attività farmacologica

Increased Gal‑3BP plasma levels in hospitalized patients infected with SARS‑CoV‑2

Coronavirus disease 2019 (COVID-19) has quickly turned into a health, fnancial and societal problem globally. The complex pathogenesis of severe acute respiratory syndrome coronavirus centers on the unpredictable clinical progression of the disease, which may evolve abruptly and results in critical and life-threatening clinical complications. Efective laboratory biomarkers that can classify patients according to risk of progression to severe disease are essential for ensuring timely treatment. Gal-3BP is a human secreted protein with innate immune functions, which is upregulated in viral infections, promotes infammation and has been shown to induce IL-6 expression. In this study, Gal-3BP plasma levels were measured retrospectively in a cohort of 84 hospitalized COVID-19 patients. These were classifed as having either “non-severe” or “severe” disease. Compared to healthy controls, Gal-3BP plasma levels were markedly increased in COVID-19 patients (P&lt; 0.0001). Moreover, the levels were higher in severe than in non-severe patients (P &lt; 0.05). As expected, patients with severe disease had plasma levels of IL-6 higher than patients with non-severe disease (P &lt; 0.01). In non-severe disease patients, Gal-3BP levels collected at a late stage (13.3 + 5.7 days after the frst positive PCR result) were signifcantly lower than those collected at an early stage (4.2 + 2.9 days form the frst positive PCR result). Larger prospective analyses are needed to strength our understanding of the prognostic utility of Gal-3BP in COVID-19 patients

ErbB-3 activation by NRG-1β sustains growth and promotes vemurafenib resistance in BRAF-V600E colon cancer stem cells (CSCs)

Approximately 5-10% of metastatic colorectal cancers harbor a BRAF-V600E mutation, which is correlated with resistance to EGFR-targeted therapies and worse clinical outcome. Vice versa, targeted inhibition of BRAF-V600E with the selective inhibitor PLX 4032 (Vemurafenib) is severely limited due to feedback re-activation of EGFR in these tumors. Mounting evidence indicates that upregulation of the ErbB-3 signaling axis may occur in response to several targeted therapeutics, including Vemurafenib, and NRG-1β-dependent re-activation of the PI3K/AKT survival pathway has been associated with therapy resistance. Here we show that colon CSCs express, next to EGFR and ErbB-2, also significant amounts of ErbB-3 on their membrane. This expression is functional as NRG-1β strongly induces AKT/PKB and ERK phosphorylation, cell proliferation, clonogenic growth and promotes resistance to Vemurafenib in BRAF-V600E mutant colon CSCs. This resistance was completely dependent on ErbB-3 expression, as evidenced by knockdown of ErbB-3. More importantly, resistance could be alleviated with therapeutic antibody blocking ErbB-3 activation, which impaired NRG-1β-driven AKT/PKB and ERK activation, clonogenic growth in vitro and tumor growth in xenograft models. In conclusion, our findings suggest that targeting ErbB-3 receptors could represent an effective therapeutic approach in BRAF-V600E mutant colon cancer