Growth Factor Modulation Research Articles

Chondroitin sulfate n-acetylgalactosaminyltransferase-2 (ChGn-2) plays a significant role in cardiac remodeling and heart failure following pressure overload

Abstract Background Cardiac extracellular matrix (ECM) is critically involved in cardiac homeostasis by providing mechanical support as well as modulating growth factor signaling. Cardiac ECM dysregulation has been shown in heart failure pathogenesis, and accumulation of chondroitin sulfate glycosaminoglycans (CS-GAGs) was previously shown to exacerbate heart failure by augmenting inflammation and fibrosis at the chronic phase. However, it remains unclear whether and the mechanism by which CS-GAGs cause cardiac dysfunction, especially at the acute phase. Purpose The purpose of this study is to elucidate the role of CS-GAGs in heart failure. Methods In this study, we analyzed the role of CS-GAGs in heart failure using mice with target deletion of chondroitin sulfate N-acetylgalactosaminyltransferase (ChGn)-2 that elongates CS chains of GAGs. Heart failure was induced by transverse aortic constriction (TAC) in mice. Since cardiac fibroblasts (CFs) are the primary cells for ECM production in the heart, we explored the role of CF-derived ECM in cardiomyocyte apoptosis. CFs were given stretch stimuli that mimic pressure overload conditions. Results Significant CS-GAGs accumulation was detected in the heart of WT mice after TAC, which was substantially reduced in the heart of ChGn2−/− mice. Unexpectedly, loss of ChGn-2 deteriorated left ventricular systolic dysfunction accompanied by augmented cardiac hypertrophy and increased cardiomyocyte apoptosis. Stretch stimuli increased ChGn-2 expression and enhanced GAG production in CFs. Interestingly, only conditioned medium (CM) derived from stretched CFs showed protective effects on cardiomyocyte death induced by doxorubicin. Degradation of CS-GAGs in CFs-derived CM by using Chondroitinase ABC abolished its cardioprotective effect. Further experiments revealed that this cardioprotective effect is at least partially through CS-GAGs-derived PI3K/AKT pathway activation via CD44. Conclusion Our data revealed that CF-derived GAGs protect cardiomyocytes from death in the acute phase of heart failure due to pressure overload; thus, insufficient GAGs production caused by loss of ChGn-2 exacerbated heart failure. Funding Acknowledgement Type of funding sources: None.

Use of Intravesical Injections of Platelet-Rich Plasma for the Treatment of Bladder Pain Syndrome: A Comprehensive Literature Review.

Background: Bladder pain syndrome/interstitial cystitis (BPS/IC) or primary bladder pain syndrome (PBPS) is a complex and poorly understood condition. This comprehensive review aimed to discuss the potential application of platelet-rich plasma (PRP) in the treatment of BPS/IC. The pathophysiology of BPS/IC is characterized by urothelial damage that triggers a chain of events leading to chronic inflammation and other conditions. Frequently, in subjects affected by BPS/IC, recurrent urinary tract infection (rUTI) is associated with difficult therapeutic management. For these reasons, many oral and intravesical treatments (e.g., antibiotic therapy and intravesical anesthetic instillations) have been proposed to alleviate the symptoms of IC/BPS. However, the limitation of these treatments is the short duration of improvement. The purpose of this review is to analyze the efficacy of intravesical PRP injections in subjects with PBS/IC and to try to understand the potential therapeutic effects on the pathophysiology of this disease. Methods: A nonsystematic literature search using Pubmed, EMBASE, Scopus, Web of Science, Medline was performed from January 2000 to August 2021. The following terms were combined to capture relevant publications: “platelet-rich plasma”, “interstitial cystitis”, “PRP”, “bladder pain syndrome”, and “painful bladder syndrome”. Results: After exclusion of non-pertinent studies/articles, we have analyzed 5 studies. In detail, 2 articles concerned preclinical studies in which animal models were used. The authors showed an improvement in the histological pattern with less bleeding in treated subjects, a lower presence of inflammatory cytokines and an increase in the mitotic index of urothelial cells in animals treated with intravesical PRP. In the three prospective clinical trials analyzed, patients with BPS/IC who underwent monthly intravesical PRP injections were found to have a statistically significant improvement in symptoms with modulation of growth factors and inflammatory proteins. Conclusions: New evidence suggests that treatment with intravesical PRP could improve urothelial regeneration and reduces chronic inflammation in BPS/IC, modifying the clinical history of its pathology.

Pre-operative Lab Values and Wound Healing following Facial Trauma Surgery: Which Values Predict Postoperative Complications?

Statement of the problemCan pre-operative lab values be used to predict wound healing in maxillofacial trauma patients? Materials and methodsAn IRB-approved (#4699) retrospective cohort study of patients with maxillofacial trauma that required surgical repair from 2014 to 2020 was performed. Patients treated by OMSs at a single Level I Trauma Center (University of Tennessee Medical Center) were included in the study. The primary predictor variables were preoperative lab values, including serum albumin, WBC, ANC, and lymphocyte count. Complications related to surgical reconstruction of facial injuries represented the primary outcome variable. Methods of data analysisIndependent T-test analysis was completed to assess for significant associations between these objective measures in relation to postoperative complications. A Chi-square test was performed to assess for differences in complication rates based on patient sex. ResultsThe patient cohort included 57 individuals; of whom, 42 were men. There was no difference in complication rates between men and women (P = .47). The average age of the patients at surgery was 38 years. Of these 57 patients, 15 had complications ranging from surgical site infection to prolonged trigeminal nerve hypothesia. When controlling for all other variables, ANC:lymphocyte ratio and number of procedures were both approaching statistical significance P = .097 and .058, respectively. Subjects experiencing complications were found to have an average ANC:lymphocyte ratio of 11.645, while those who had no complications had an average ANC:lympocyte ratio of 7.759. Pre-operative albumin was not found to be statistically significant in maxillofacial trauma patients with a P = .522. Patients without complication had an average albumin of 4.011, and those with complication average an albumin level of 4.143. Outcomes dataSee attached tables. ConclusionLymphocytes are recognized as critical regulators to wound healing via modulation and regulation of growth factors and cytokines.1 Often observed, but rarely characterized, differences in preoperative lymphocyte to neutrophil ratios seem to relate a risk of complication to the patient. Additionally, recent literature has reflected an association between poor nutritional status, with albumin levels used as a surrogate marker, and postoperative complications. This correlation has been noted in oncologic procedures of the head and neck in recent years.2 The authors' study demonstrated that among those individuals meeting the inclusion criteria, there may be an association between higher ANC:lymphocyte levels and postoperative complication rates in facial trauma patients. However, there was no relation between preoperative albumin levels and complications in patients. Further research and data collection are being completed in an effort to increase the power of these results.

Chitosan-Coated PLGA Nanoparticles Encapsulating Triamcinolone Acetonide as a Potential Candidate for Sustained Ocular Drug Delivery.

The current treatment for the acquired retinal vasculopathies involves lifelong repeated intravitreal injections of either anti-vascular endothelial growth factor (VEGF) therapy or modulation of inflammation with steroids. Consequently, any treatment modification that decreases this treatment burden for patients and doctors alike would be a welcome intervention. To that end, this research aims to develop a topically applied nanoparticulate system encapsulating a corticosteroid for extended drug release. Poly (lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) supports the controlled release of the encapsulated drug, while surface modification of these NPs with chitosan might prolong the mucoadhesion ability leading to improved bioavailability of the drug. Triamcinolone acetonide (TA)-loaded chitosan-coated PLGA NPs were fabricated using the oil-in-water emulsion technique. The optimized surface-modified NPs obtained using Box-Behnken response surface statistical design were reproducible with a particle diameter of 334 ± 67.95 to 386 ± 15.14 nm and PDI between 0.09 and 0.15. These NPs encapsulated 55–57% of TA and displayed a controlled release of the drug reaching a plateau in 27 h. Fourier-transform infrared spectroscopic (FTIR) analysis demonstrated characteristic peaks for chitosan (C-H, CONH2 and C-O at 2935, 1631 and 1087 cm−1, respectively) in chitosan-coated PLGA NPs. This result data, coupled with positive zeta potential values (ranged between +26 and +33 mV), suggests the successful coating of chitosan onto PLGA NPs. Upon coating of the NPs, the thermal stability of the drug, polymer, surfactant and PLGA NPs have been enhanced. The characteristics of the surface-modified NPs supports their use as potential candidates for topical ocular drug delivery for acquired retinal vasculopathies.

Pentosan polysulfate sodium prevents functional decline in chikungunya infected mice by modulating growth factor signalling and lymphocyte activation.

Chikungunya virus (CHIKV) is an arthropod-borne virus that causes large outbreaks world-wide leaving millions of people with severe and debilitating arthritis. Interestingly, clinical presentation of CHIKV arthritides have many overlapping features with rheumatoid arthritis including cellular and cytokine pathways that lead to disease development and progression. Currently, there are no specific treatments or vaccines available to treat CHIKV infections therefore advocating the need for the development of novel therapeutic strategies to treat CHIKV rheumatic disease. Herein, we provide an in-depth analysis of an efficacious new treatment for CHIKV arthritis with a semi-synthetic sulphated polysaccharide, Pentosan Polysulfate Sodium (PPS). Mice treated with PPS showed significant functional improvement as measured by grip strength and a reduction in hind limb foot swelling. Histological analysis of the affected joint showed local inflammation was reduced as seen by a decreased number of infiltrating immune cells. Additionally, joint cartilage was protected as demonstrated by increased proteoglycan staining. Using a multiplex-immunoassay system, we also showed that at peak disease, PPS treatment led to a systemic reduction of the chemokines CXCL1, CCL2 (MCP-1), CCL7 (MCP-3) and CCL12 (MCP-5) which may be associated with the reduction in cellular infiltrates. Further characterisation of the local effect of PPS in its action to reduce joint and muscle inflammation was performed using NanoString™ technology. Results showed that PPS altered the local expression of key functional genes characterised for their involvement in growth factor signalling and lymphocyte activation. Overall, this study shows that PPS is a promising treatment for alphaviral arthritis by reducing inflammation and protecting joint integrity.

Lithium as a Neuroprotective Agent for Bipolar Disorder: An Overview.

Lithium (Li+) is a first option treatment for adult acute episodes of Bipolar Disorder (BD) and for the prophylaxis of new depressed or manic episodes. It is also the preferred choice as maintenance treatment. Numerous studies have shown morphological abnormalities in the brains of BD patients, suggesting that this highly heritable disorder may exhibit progressive and deleterious changes in brain structure. Since treatment with Li+ ameliorates these abnormalities, it has been postulated that Li+ is a neuroprotective agent in the same way atypical antipsychotics are neuroprotective in patients diagnosed with schizophrenia spectrum disorders. Li+'s neuroprotective properties are related to its modulation of nerve growth factors, inflammation, mitochondrial function, oxidative stress, and programmed cell death mechanisms such as autophagy and apoptosis. Notwithstanding, it is not known whether Li+-induced neuroprotection is related to the inhibition of its putative molecular targets in a BD episode: the enzymes inositol-monophosphatase, (IMPase), glycogen-synthase-kinase 3β (GSK3), and Protein kinase C (PKC). Furthermore, it is uncertain whether these neuroprotective mechanisms are correlated with Li+'s clinical efficacy in maintaining mood stability. It is expected that in a nearby future, precision medicine approaches will improve diagnosis and expand treatment options. This will certainly contribute to ameliorating the medical and economic burden created by this devastating mood disorder.

Study of cancer cell cytotoxicity, internalization and modulation of growth factors induced by transferrin-conjugated formulations of metallodrug-functionalized mesoporous silica nanoparticles

Nano-sized materials have shown to have very high potential in anticancer therapy by an adequate tuning of their functionalization and physico-chemical properties. This study is focused on the synthesis and characterization of mesoporous silica nanoparticles (MSN) functionalized with a titanium(IV) or an organotin(IV) compound (therapeutic agents), fluorescein isothiocyanate (image agent) and transferrin (targeting molecule). The analysis of the biological activity of the metallodrug-functionalized systems with and without transferrin has been evaluated and the cell internalization with respect to the presence of the protein has been assessed. The biological results show, as expected, that Sn-based materials are more active than the Ti-based systems with some of the tin-functionalized nanoparticles being almost 50 times more active than carboplatin. In contrast, the cellular uptake seems to be higher in Ti-based materials, which take advantage of the stronger interaction with transferrin to internalize the cells with more effectivity. Finally, a study of vascular endothelial growth factor A (VEGF-A), human fibroblast growth factor 2 (FGF-2) and nuclear factor κβ transcription factor (NF-κβ) show that, specially the Sn-based MSNs, were able to modulate these factors in A2780 cells showing anti-angiogenic effects through VEGF-A and FGF-2, probably due to interaction of the materials with transferrin. • MSN have been functionalized with Sn or Ti metallodrugs and FITC and formulated with transferrin. • Transferrin assists in the internalization of all the functionalized mesoporous nanoparticles. • Transferrin does not increase the cytotoxicity of the functionalized mesoporous systems. • Transferrin-functionalized MSN materials modulate and downregulate growth and transcription factors.

A robust platform for high-throughput screening of therapeutic strategies for acute and chronic spinal cord injury

SummaryAstrocytes and microglia are critical regulators of inflammatory cascade after spinal cord injury (SCI). Existing glial in vitro studies do not replicate inflammatory phases associated with SCI. Here, we report an in vitro model of mixed glial culture where inflammation is induced by the administration of pro-inflammatory cytokines (tumor necrosis factor-α, interleukin-1β, and interleukin-6) to promote pathologically relevant “acute” and “chronic” inflammatory phases. We observed SCI relevant differential modulation of inflammatory pathways, cytokines, chemokines, and growth factors over 21 days. Mitochondrial dysfunction was associated with a cytokine combination treatment. Highly expressed cytokine induced neutrophil chemoattractant (CINC-3) chemokine was used as a biomarker to establish an enzyme-linked immunosorbent assay-based high-throughput screening (HTS) platform. We screened a 786-compound drug library to demonstrate the efficacy of the HTS platform. The developed model is robust and will facilitate in vitro screening of anti-reactive glial therapeutics for the treatment of SCI.

RIPK3 modulates growth factor receptor expression in endothelial cells to support angiogenesis.

Receptor-interacting protein kinase 3 (RIPK3) is a multifunctional intracellular protein that was first recognized as an important component of the necroptosis programmed cell death pathway. RIPK3 is also highly expressed in non-necroptotic murine embryonic endothelial cells (ECs) during vascular development, indicating its potential contribution to angiogenesis. To test this hypothesis, we generated mice lacking endothelial RIPK3 and found non-lethal embryonic and perinatal angiogenesis defects in multiple vascular beds. Our in vitro data indicate that RIPK3 supports angiogenesis by regulating growth factor receptor degradation in ECs. We found that RIPK3 interacted with the membrane trafficking protein myoferlin to sustain expression of vascular endothelial growth factor receptor 2 (VEGFR2) in cultured ECs following vascular endothelial growth factor A (VEGFA) stimulation. Restoration of myoferlin, which was diminished after RIPK3 knockdown, rescued decreased VEGFR2 expression and vascular sprouting in RIPK3-deficient ECs after VEGFA treatment. In addition, we found that RIPK3 modulated expression of genes involved in endothelial identity by inhibiting ERK signaling independently of growth factor receptor turnover. Altogether, our data reveal unexpected non-necroptotic roles for RIPK3 in ECs and evidence that RIPK3 promotes developmental angiogenesis in vivo.

Structural studies of reelin N-terminal region provides insights into a unique structural arrangement and functional multimerization.

The large, secreted glycoprotein reelin regulates embryonic brain development as well as adult brain functions. Although reelin binds to its receptors via its central part, the N-terminal region directs multimer formation and is critical for efficient signal transduction. In fact, the inhibitory antibody CR-50 interacts with the N-terminal region and prevents higher-order multimerization and signalling. Reelin is a multidomain protein in which the central part is composed of eight characteristic repeats, named reelin repeats, each of which is further divided by insertion of a epidermal growth factor (EGF) module into two subrepeats. In contrast, the N-terminal region shows unique 'irregular' domain architecture since it comprises three consecutive subrepeats without the intervening EGF module. Here, we determined the crystal structure of the murine reelin fragment named RX-R1 including the irregular region and the first reelin repeat at 2.0-Å resolution. The overall structure of RX-R1 has a branched Y-shaped form. Interestingly, two incomplete subrepeats cooperatively form one entire subrepeat structure, though an additional subrepeat is inserted between them. We further reveal that Arg335 of RX-R1 is crucial for binding CR-50. A possible self-association mechanism via the N-terminal region is proposed based on our results.

Association of steroid use with survival in solid tumours.

Steroids are commonly used in patients with solid tumours for supportive therapy. In other cases, they are an essential part of cancer treatment such as prostate cancer. Some preclinical observations lead to the notion that glucocorticoids may modulate growth factors' pathways and may induce the progression of cancers. Glucocorticoids are associated with several side-effects on many organ systems (e.g. serious infections, diabetes, sepsisand thrombosis). We have performed a systematic review and meta-analysis to evaluate the outcome of cancer patients that assume or not steroids. Published articles that evaluated survival associated with steroids use in cancer patients from inception to June 2020 were identified by searching the PubMed, EMBASEand Cochrane Library databases. The primary outcome of interest was the risk of death, and the secondary end-point was the risk of progression in steroid versus non-steroid users. Seventy-six studies were in quantitative synthesis for a total of 83,614 patients. Use of steroids was associated with a reduced survival (hazard ratios (HR)=1.18, 95% confidence interval (CI): 1.1-1.26; P<.01). Progression-free survival was also decreased in steroid versus non-steroid users (HR=1.13, 95% CI: 1.01-1.26; P=.03). In patients with lung cancer, advanced disease and supportive care indications were settings where the use of steroids increased the risk of death. In patients with advanced cancers, use of steroids should be reduced and, at best, avoided because it may reduce survival, in particular, for patients with lung cancer and for palliative/supportive care purposes.

Mechanisms underlying the wound healing and tissue regeneration properties of Chenopodium album.

In the current study, aerial parts (leaves, stem and shoots) of C. album were extracted with methanol and subjected to phytochemical and HPLC analysis. Agar well diffusion method was used for anti-bacterial activity against Gram-negative strains Escherichia coli, Salmonella typhi, Klebsiella, Pseudomonas aeruginosa and Gram-positive Bacillus cereus, Staphylococcus aureus, Methicillin-resistant Staphylococcus aureus. Burn was induced through flame heated metal rod on mice. C. album ointment (2% w/w), Vaseline (vehicle) and silver sulfadiazine (standard) were topically applied thrice daily for 15days. Wound area was measured on day 0, 5, 10 and 15. On last day, the wound tissues were excised and subjected to histopathological, quantitative PCR and immunohistochemical analysis. Phenols, alkaloids, phytosterols, tannins, saponins, flavonoids, carbohydrates and glycosides were detected in phytochemical analysis. HPLC chromatogram displayed peaks for rutin, quercetin, ascorbic acid, gallic acid and various other phyto-constituents. The extract exhibited zone of inhibition in millimeter (mm) against E. coli (12.3 ± 0.57), S. typhyi (14.6 ± 1.52), Klebsiella (11.8 ± 0.76), P. aeruginosa (12.3 ± 0.57), B. cereus (12.5 ± 1.29), S. aureus (18.3 ± 2.08), and MRSA (11.8 ± 0.76). The wound area in C. album group was significantly (60%) reduced as compared to vehicle group (11%). Histological analysis showed complete re-epithelialization and fine tissue in extract treated group. qPCR data revealed up-regulation of EGF, PDGF and TGF-β1 genes in extract treated group. Similarly, immunohistochemistry results confirmed heightened EGFR expression in extract treated group. Our findings suggest that C. album can promote wound healing and tissue regeneration through control of burns related infection and modulation of growth factors and its receptors.

1944P Hyperglycemia increased nivolumab-induced cardiotoxicity, enhanced immunoresistance of ERɑ+, PR+, HER2- breast cancer cells modulating growth factors and NLRP3 expression

Type 2 diabetes and metabolic syndrome are negative prognostic factors in breast cancer patients. Immune checkpoint inhibitors (ICIs) have revolutionized cancer treatment, achieving unprecedented efficacy in multiple malignancies. We aimed to study if hyperglycemia could enhance ICI-induced cardiotoxicity in cardiomyocites and immunoresistance in human breast cancer cells. Finally, we evaluated if the treatment with an SGLT-2 inhibitor (empagliflozin) could revert these effects during hyperglycemic condition. Human cardiomyocytes (HL-1 cells) and PD-1+ ERɑ+, PR+, HER2- breast cancer cells (MCF-7 cell line) were exposed to nivolumab (100 nM) at high glucose (25 mM) low glucose (5.5 mM) for 72 h. After the incubation period, we performed the following tests: determination of cell viability, through analysis of mitochondrial dehydrogenase activity; NLRP3, p65/NF-kB and leukotrienes expression through ELISA method. Nivolumab-induced cardiotoxicity was enhanced by 2,7-fold during exposure to 25 mM glucose (High Glucose; HG) compared to 5.5 mM glucose (Low Glucose; LG) in human adult cardiomyocytes. Moreover, IC50 value of nivolumab against MCF7-cells increased significantly under HG vs LG (P<0.01). Moreover, during high glucose condition, cardiomyocytes and human breast cancer cells exposed to nivolumab, increases the expression of NLRP3, p65/NF-kB, leukotrienes and cytokines. Notably, hyperglycemia increases significantly the intracellular calcium (iCa2+) content in cardiomyocytes during incubation with nivolumab, probably increasing the metabolic susceptibility to damages induced by ICI. Shifting from HG to LG, as well as the administration of 50 nM empagliflozin reduced the magnitude of cardiotoxic effects, indicating cardioprotective and immuno-enhancing properties. To our knowledge these results are the first evidence that hyperglycemia exacerbates ICI-induced cardiotoxicity and immunoresistace and set the stage to preclinical and clinical trials aimed to decrease glucose through dietary or lifestyle changes or through new hypoglycemic drugs (gliflozins).

Late-onset hypogonadism: Reductio ad absurdum of the cardiovascular risk-benefit of testosterone replacement therapy.

Low testosterone (T) level is considered a marker of poor cardiovascular health. Ten years ago, the Testosterone in Older Men with Mobility Limitations (TOM) trial was discontinued due to a higher number of adverse events in men receiving T compared with placebo. Since then, several studies have investigated the risks of T replacement therapy (TRT) in late-onset hypogonadism (LOH). To review the mechanism by which TRT could damage the cardiovascular system. Comprehensive literature search of recent clinical and experimental studies. The mechanisms of T-mediated coronary vasodilation were reviewed with emphasis on calcium-activated and ATP-sensitive potassium ion channels. We showed how T regulates endothelial nitric oxide synthase (eNOS) and phosphoinositide 3-kinase/protein kinase B/eNOS signaling pathways in vessel walls and its direct effects on cardiomyocytes via β1-adrenergic and ryanodine receptors and provided data on myocardial infarction and heart failure. Vascular smooth muscle senescence could be explained by the modulation of growth factors, matrix metalloproteinase-2, and angiotensin II by T. Furthermore, leukocyte trafficking, facilitated by changes in TNF-α, could explain some of the effects of T on atheromatous plaques. Conflicting data on prothrombotic risk linked to platelet aggregation inhibition via NO-triggered arachidonate synthesis or increased aggregability due to enhanced thromboxane A in human platelets provide evidence regarding the hypotheses on plaque maturation and rupture risk. The effects of T on cardiac electrophysiology and oxygen delivery were also reviewed. The effects of TRT on the cardiovascular system are complex. Although molecular studies suggest a potential benefit, several clinical observations reveal neutral or occasionally detrimental effects, mostly due to confounding factors. Attempts to demonstrate that TRT damages the cardiovascular system via systematic analysis of the putative mechanisms led to the contradiction of the initial hypothesis. Current evidence indicates that TRT is safe once other comorbidities are addressed.

Abstract PHB04: Predictive modeling, applied to genetically engineered mouse models of breast or lung cancer, provides insights into major oncogenic pathways

Abstract The H2020 project CanPathPro is building and validating a computational predictive modelling platform applied to cancer. To this end, we develop and refine bioinformatic and experimental tools, utilized in generation and evaluation of systems biology modeling predictions. The presented work employs the following technologies and methodologies: biologic systems representing 3 levels of biologic complexity (genetically engineered mouse models—GEMMs—of breast or lung cancer, organoids and cell lines derived thereof); next-generation sequencing and SWATH-based phospho/proteomics; and two large-scale computational mechanistic models. The highly defined biologic systems are used (i) to activate selected oncogenic stimuli that modulate pathway components in a systematic manner; (ii) to characterize the signaling changes occurring during cancer development—thus generating temporally resolved datasets for model training; and (iii) to validate, in vitro and in vivo, the modeling predictions. The mechanistic models, based on ordinary differential equations, enable prediction of phenotypes and drug response in mouse or human. Model parameters are defined using project-derived experimental data, either via parameter estimation strategies or via selection of parameter distributions by a Monte Carlo approach. For simulations, the models are initialized with transcriptome data, either from GEMM-derived cell lines grown under variable conditions or from GEMM-derived neoplastic and tumor tissue, representing lesion progression. The models also integrate the relevant mutations. Results, obtained by iterative rounds of in silico predictions and in vitro validation, include in silico identification (i) of the activation status of oncogenic pathways in GEMMs, organoids, and cell lines; (ii) of the signaling changes induced by the in vitro growth conditions (e.g., growth factor modulation, drug treatments) and by the mutational profile of cell lines/organoids, or by the mutational profile and lesion stage of each GEMM; and (iii) of the drug response of cell lines and GEMMs. In conclusion, this work encompasses a highly integrative systems biology approach generating and validating new hypotheses on cancer pathways signaling and crosstalk, identifying new signal flow, and suggests new ways to interfere with tumor growth. This abstract is also being presented as Poster B49. Citation Format: Julio Banga, Lucien Frappart, Jan Hasenauer, Yann Herault, Jos Jonkers, David Koubi, Bodo Lange, Glenn Terje Lines, Aspasia Plouidou, Oliver Rinner. Predictive modeling, applied to genetically engineered mouse models of breast or lung cancer, provides insights into major oncogenic pathways [abstract]. In: Proceedings of the AACR Special Conference on the Evolving Landscape of Cancer Modeling; 2020 Mar 2-5; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2020;80(11 Suppl):Abstract nr PHB04.

Quercetin loaded chitosan tripolyphosphate nanoparticles accelerated cutaneous wound healing in Wistar rats

Previous studies have shown that quercetin on topical application improved cutaneous wound healing in rats, but hydrophobic nature and less skin penetration limits its potential as topical healing agent. Therefore, present study was planned to investigate wound healing potential of chitosan based quercetin nanoparticles. Quercetin loaded nanoparticles were synthesized by ionic gelation method and characterized by various standard techniques. A 2 × 2 cm2 square shaped wound was created on the thoraco-lumbar part of rats. Wounded rats were divided into 6 groups namely, gel (20%), blank nanoparticle (0.16%), bulk quercetin (0.3%), quercetin nanoparticles (0.03%), quercetin nanoparticles (0.1%) and quercetin nanoparticles (0.3%) treated groups. Different formulations of quercetin nanoparticles were applied on the wounds for the duration of study and healing tissues were collected on 7th, 14th and 21st day to study various parameters. Quercetin loaded nanoparticles were 361.16 ± 9.72 nm size and spherical in shape. We observed quercetin nanoparticles (0.03%) treatment caused marked reduction in the tumor necrosis factor-alpha, whereas expressions of interleukin 10, vascular endothelial growth factor and transforming growth factor beta1 was increased significantly with treatment. The granulation tissue of quercetin nanoparticles (0.03%) treated group showed better quality healing and maturity as supported by the increased blood vessels density, decreased inflammatory cells, increased number of myofibroblasts, deposition and arrangement of collagen fibers and re-epithelialization. In conclusion, quercetin nanoparticles (0.03%) treatment significantly improved wound healing by modulation of cytokines and growth factors involved in inflammatory and proliferative phases of wound healing.

Deriving Immune Modulating Drugs from Viruses-A New Class of Biologics.

Viruses are widely used as a platform for the production of therapeutics. Vaccines containing live, dead and components of viruses, gene therapy vectors and oncolytic viruses are key examples of clinically-approved therapeutic uses for viruses. Despite this, the use of virus-derived proteins as natural sources for immune modulators remains in the early stages of development. Viruses have evolved complex, highly effective approaches for immune evasion. Originally developed for protection against host immune responses, viral immune-modulating proteins are extraordinarily potent, often functioning at picomolar concentrations. These complex viral intracellular parasites have “performed the R&D”, developing highly effective immune evasive strategies over millions of years. These proteins provide a new and natural source for immune-modulating therapeutics, similar in many ways to penicillin being developed from mold or streptokinase from bacteria. Virus-derived serine proteinase inhibitors (serpins), chemokine modulating proteins, complement control, inflammasome inhibition, growth factors (e.g., viral vascular endothelial growth factor) and cytokine mimics (e.g., viral interleukin 10) and/or inhibitors (e.g., tumor necrosis factor) have now been identified that target central immunological response pathways. We review here current development of virus-derived immune-modulating biologics with efficacy demonstrated in pre-clinical or clinical studies, focusing on pox and herpesviruses-derived immune-modulating therapeutics.

Polychlorinated dibezno-p-dioxins and dibenzofurans. Documentation of proposed values of occupational exposure limits (OELs)

Polychlorinated dibezno-p-dioxins (PCDD) and dibenzofurans (PCDF) are compounds with similar structure, physicochemical and toxicological properties. They are not used commercially, they are formed as by-products during certain industrial processes, combustion, failures, etc. LD50 values (0.002-300 mg / kg) depend on the species of animal tested and the chemical structure of the compound. On chronic toxicity, the information available mainly relates to 2,3,7,8-TCDD and 2,3,4,7,8-PeCDF. Potential routes of human exposure are: the digestive system, lungs and skin. These compounds are accumulated mainly in the liver and adipose tissue. Their polar metabolites may undergo conjugation with glucuronic acid and glutathione. The main routes of excretion are bile and feces. In mammals, PCDD / F are also eliminated in breast milk. The results of mutagenicity and genotoxicity tests of PCDD (mainly 2,3,7,8-TCDD) and PCDF and their effects on fertility and reproduction are inconsistent. Among PCDD and PCDF, the compound that most strongly affects fertility, reproduction and fetal development is 2,3,7,8-TCDD. Epidemiological studies are the basis for assessing the carcinogenic potential of dioxins (including 2,3,7,8-TCDD) and furans in humans. Cohorts include those professionally exposed to chlorophenols, phenoxyacetic herbicides and a mixture of polychlorinated dibenzodioxins and furans. PCDD / F have a common mechanism of toxic action associated with the Ah receptor. PCDD / F are considered to be inducers of several enzymes (e.g. CYP1A) and modulators of hormones and growth factors. CYP1A1 activity is one of the most sensitive indicators of exposure to 2,3,7,8-TCDD. Adenocarcinomas and hepatocellular carcinomas as well as bile ducts have been found in rats and mice after 2,3,7,8-TCDD. Tumor changes have also been observed in other organs. NTP studies also showed carcinogenic effects 2,3,4,7,8-PeCDF. According to IARC, sufficient evidence of a carcinogenic effect on humans exists only for 2,3,7,8-TCDD (CAS: 1746-01-6) and 2,3,4,7,8 PeCDF (CAS: 57117-31-4). Other PCDD / F cannot be classified as carcinogenic to humans. The basis for determining the MAC value for the mixture of PCDD and PCDF was the results of the assessment of the risk of developing additional liver cancer in people exposed in the work environment for 2,3,7,8-TCDD in 2017. This risk was estimated at 1 · 10-4 for 40 years of exposure to the compound at a concentration of 18 pg / m3. In the case of combined exposure, the content of polychlorinated dibenzo-p-dioxins and furans in the tested samples, as well as their maximum acceptable levels are expressed in the form of the so-called toxicity equivalent (TEQ). For the PCDD and PCDF mixture, we propose the value of 18 pg WHO2006-TEQ / m3. The result expressed as pg WHO-TEQ / m3 is not a de facto concentration, but a determination of the toxicity of the mixture of dioxin and furan congeners contained in the sample in relation to TCDD. This article discusses the problems of occupational safety and health, which are covered by health sciences and environmental engineering.

Quercetin accelerated cutaneous wound healing in rats by modulation of different cytokines and growth factors

Quercetin on wounds could be favorable for healing based on its variety of biological effects. Eighty wounded rats were divided into four groups i.e. dimethyl sulfoxide, 0.03% quercetin, 0.1% quercetin, and 0.3% quercetin-treated. Different treatments were topically applied for 20 days. Quercetin (0.3%) caused the fastest wound closure and markedly improved the oxidative stress. Quercetin treatment increased the expressions of IL-10, VEGF, TGF-β1, CD31, α-SMA, PCNA, and GAP-43, and decreased the expressions of TNF-α. Early infiltration of inflammatory cells and formation of good quality granulation tissue dominated by fibroblast proliferation, angiogenesis, and collagen deposition in quercetin treated groups was also evident. All these effects were more pronounced at 0.3% quercetin concentration. The earliest regeneration of epithelial layer was also observed in 0.3% quercetin-treated wounds. In conclusion, 0.3% quercetin accelerates wound healing efficiently by modulating antioxidant system of wound, cytokines, growth factors, other proteins and cells involved in healing.

A Fyn biosensor reveals pulsatile, spatially localized kinase activity and signaling crosstalk in live mammalian cells.

Cell behavior is controlled through spatio-temporally localized protein activity. Despite unique and often contradictory roles played by Src-family-kinases (SFKs) in regulating cell physiology, activity patterns of individual SFKs have remained elusive. Here, we report a biosensor for specifically visualizing active conformation of SFK-Fyn in live cells. We deployed combinatorial library screening to isolate a binding-protein (F29) targeting activated Fyn. Nuclear-magnetic-resonance (NMR) analysis provides the structural basis of F29 specificity for Fyn over homologous SFKs. Using F29, we engineered a sensitive, minimally-perturbing fluorescence-resonance-energy-transfer (FRET) biosensor (FynSensor) that reveals cellular Fyn activity to be spatially localized, pulsatile and sensitive to adhesion/integrin signaling. Strikingly, growth factor stimulation further enhanced Fyn activity in pre-activated intracellular zones. However, inhibition of focal-adhesion-kinase activity not only attenuates Fyn activity, but abolishes growth-factor modulation. FynSensor imaging uncovers spatially organized, sensitized signaling clusters, direct crosstalk between integrin and growth-factor-signaling, and clarifies how compartmentalized Src-kinase activity may drive cell fate.