Targeting Growth Factor Research Articles

Comparison of Corneal Epitheliotrophic Factors of Undiluted Autologous Platelet-Rich Plasma and Autologous Serum Eye Drops for Dry Eye Disease.

The purpose of this study was to compare the growth factor concentrations in undiluted autologous platelet-rich plasma (APRP) and autologous serum (AS) eye drops. This was a single-center, prospective trial conducted in a tertiary university hospital in Bangkok, Thailand. Ninety-six patients with moderate-to-severe dry eye disease, who were randomly assigned to receive either 100% APRP or 100% AS, were enrolled in the study. Primary outcome measures were the concentrations of epitheliotrophic factors, including epithelial growth factor (EGF), basic fibroblast growth factor (bFGF), hepatocyte growth factor (HGF), β-nerve growth factor (β-NGF), platelet-derived growth factor (PDGF)-AA, PDGF-BB, transforming growth factor (TGF)-α, TGF-β1, and vascular endothelial growth factor (VEGF) in 100% APRP and 100% AS. Secondary outcome measures were the correlations between baseline patient characteristics and each growth factor concentration. Undiluted APRP contained more EGF, bFGF, and β-NGF than undiluted AS (P < 0.001, P < 0.001, P = 0.018, respectively). Meanwhile, undiluted AS yielded higher concentrations of HGF, PDGF-AA, PDGF-BB, and VEGF compared with undiluted APRP (P < 0.001 all). There were no significant differences in TGF-α and TGF-β1 concentrations between the two groups. In the 100% APRP group, the platelet concentrations had positive correlations with the concentrations of EGF (P = 0.028) and VEGF (P = 0.020). In the 100% AS group, Sjögren's syndrome negatively correlated with the concentrations of PDGF-BB (P = 0.028) and VEGF (P = 0.023). Diabetes mellitus (DM) showed negative correlations with the concentrations of HGF (P = 0.001), TGF-α (P = 0.001), and VEGF (P = 0.002). With our simple preparation protocols, 100% APRP contains higher concentrations of EGF, bFGF, and β-NGF, whereas 100% AS contains higher concentrations of HGF, PDGF-AA, PDGF-BB, and VEGF. This could allow ophthalmologists to tailor treatments to individual patients by targeting growth factor supply based on their underlying pathophysiology. ClinicalTrials.gov identifier, NCT04683796.

Abstract 126: Non‐Clinical Risk Assessment of Co‐Administration of Thrombolytics and Scp776, a Targeted IGF‐1 Neuroprotectant for Stroke

Introduction/Purpose Scp776 is a first‐in‐class targeted growth factor therapeutic in development for treatment of acute ischemic stroke (AIS) patients. The therapeutic hypothesis for scp776 neuroprotection in stroke is targeted delivery of a potent growth factor (IGF‐1) alongside standard‐of‐care flow restoration will protect tissue from damage by promoting apoptosis escape. In the ARPEGGIO study of scp776 in patients undergoing endovascular thrombectomy (EVT), those receiving intravenous thrombolytics (IVT) are excluded. Before expanding testing of scp776 into the broader patient population (i.e., including IVT and IVT/EVT patients), we characterized the compatibility of scp776 with IVTs in an array of non‐clinical studies. We present tests of compatibility and propose criteria for acceptable test results to allow the safe introduction of adjuvants (e.g., neuroprotectants) to the significant population of stroke patients receiving thrombolytics. Materials/Methods The potential for interference with tPA‐mediated clot lysis by scp776 was tested in a plate‐based assay using clots derived from rat blood. The susceptibility of scp776 to proteolysis and/or inactivation by tPA or downstream effectors (e.g., plasmin) was assessed by co‐administration pharmacokinetic experiments in monkeys and a binding assay. The risk of increased bleeding in animals after co‐administration of scp776 and tPA was assessed using a mouse tail snip model. Results In the clot lysis assay, supra‐clinical concentrations of scp776 did not affect the rate or extent of clot lysis by tPA (50% Lysis Time, 40 nM tPA = 17.5±0.6 min; 40 nM tPA+100 µg/mL scp776 = 16.5±0.6 min), while the inclusion of protease inhibitors abrogated clot lysis. Scp776 remained intact, with an unchanged pharmacokinetic profile for 48 hours (~6 half‐lives) following co‐administration of 4 mg/kg scp776 with 0.9 mg/kg tPA in monkeys. The targeting function of scp776, phosphatidylserine binding, was unaffected by incubation with tPA in an in vitro binding assay (relative binding, 101%; 95% CI, 94‐109%). In the tail snip model, mice that received 5, 10, or 20 mg/kg scp776 did not have increased blood loss compared to negative controls, while mice that received 10 mg/kg tPA showed a non‐significant increase in blood loss. Mice that received 10 mg/kg tPA with 20 mg/kg scp776 showed a similar non‐significant increase in blood loss compared to negative controls. The tPA only and scp776 + tPA groups were not significantly different. Conclusion We propose a non‐clinical testing scheme for the assessment of pharmacocompatibility of novel adjuvant therapies with standard‐of‐care thrombolytics. Under this paradigm, the proposed neuroprotectant scp776 did not evidence either bidirectional drug‐drug interactions or potential synergy in dysregulation of hemostasis. Scp776 did not interfere with tPA‐mediated clot lysis in an ex vivo assay. Studies in animals and assay plates revealed that scp776 pharmacokinetics, integrity, and targeting function were not affected by tPA. Together, these results indicate that a drug‐drug interaction is unlikely in patients that receive both drugs. Co‐administration of scp776 and tPA in mice did not induce significant changes in blood loss compared to mice that received tPA only or saline. These data support future human testing of scp776 in AIS patients receiving thrombolytic therapies.

Monoclonal Antibodies and Antibody-drug Conjugates as Emerging Therapeutics for Breast Cancer Treatment.

When breast cells divide and multiply out of control, it is called breast cancer. Symptoms include lump formation in the breast, a change in the texture or color of the breast, or a discharge from the nipple. Local or systemic therapy is frequently used to treat breast cancer. Surgical and radiation procedures limited to the affected area are examples of local management. There has been significant worldwide progress in the development of monoclonal antibodies (mAbs) since 1986, when the first therapeutic mAb, Orthoclone OKT3, became commercially available. mAbs can resist the expansion of cancer cells by inducing the destruction of cellular membranes, blocking immune system inhibitors, and preventing the formation of new blood vessels. mAbs can also target growth factor receptors. Understanding the molecular pathways involved in tumor growth and its microenvironment is crucial for developing effective targeted cancer therapeutics. Due to their unique properties, mAbs have a wide range of clinical applications. Antibody-drug conjugates (ADCs) are drugs that improve the therapeutic index by combining an antigen-specific antibody with a payload. This review focuses on the therapeutic applications, mechanistic insights, characteristics, safety aspects, and adverse events of mAbs like trastuzumab, bevacizumab, pertuzumab, ertumaxomab, and atezolizumab in breast cancer treatment. The creation of novel technologies utilizing modified antibodies, such as fragments, conjugates, and multi-specific antibodies, must be a central focus of future studies. This review will help scientists working on developing mAbs to treat cancers more effectively.

First-in-human autologous CAR-T for metastatic breast cancers to target growth factor receptor MUC1*.

TPS2683 Background: MUC1 has been one of the most important therapeutic targets for solid tumor cancers for the past 20+ years. However, no MUC1 targeted therapeutic has yet succeeded or has been granted FDA approval. Although MUC1 is overexpressed on cancer cells, it is also widely expressed on normal epithelial cells. We previously reported that a MUC1 transmembrane cleavage product, MUC1* (muk 1 star), is a Class I growth factor receptor activated by ligand-induced dimerization of its truncated extra cellular domain. We generated monoclonal antibodies that recognize the specific conformation that is created when MUC1 is cleaved by specific enzymes expressed in the tumor microenvironment. 93% of human breast cancer tissues are recognized by one of these antibodies, huMNC2, which importantly does not bind to normal MUC1. We have incorporated this antibody into two CARs, wherein one bears the “1XX” mutations in CD3z that greatly increase CAR-T cell persistence, inhibit exhaustion and enable the killing of low antigen expressing cancer cells. Methods: Phase I for huMNC2-CAR44, an autologous frozen product, opened January, 2020, but as a 1st-in-human trial was paused during COVID. All patients receive 300 mg/m2 cyclophosphamide and 30 mg/m2 fludarabine for 3 days prior to CAR-T treatment. Dose escalation from 3.3x105 – 1x107 CAR+ cells/kg follows a standard 3 + 3 design. In addition to standard organ function inclusion criteria, the trial was first open to breast cancer patients whose cancer had progressed after at least 2 or 3 prior therapies while metastatic, yet there was no limit on the number of prior therapies. A recent patient biopsy had to be at least 30% positive for MUC1* in a CLIA validated IHC assay. Importantly, there were no limits on the vein-to-vein time. Dose levels 1 and 2 for huMNC2-CAR44 were completed without DLTs. Phase I for huMNC2-CAR22, which bore the 1XX mutations in CD3z to increase persistence and increase the killing of low antigen expressing cells opened September, 2023. Based on responses of initial huMNC2-CAR44 patients, inclusion criteria were amended to an Enrichment Trial Design. The number of prior therapies was limited to less than or equal to 10. Anticipated survival at the time of product infusion should be at least 3 months. Vein-to-vein time, which historically had been 15-16 days at one site and as high as 83 days at another site, would be targeted to 16- 22 days. The tumors of eligible patients need to have a MUC1* membrane positive H-score equal to or greater than 120 out of a possible 300, defined as high MUC1* positivity. Based on IHC analysis of human breast cancer TMAs, the H-score restriction would include roughly 40% of breast cancers. The increased ability of huMNC2-CAR22 to kill low antigen expressing cancer cells may support future studies of patients expressing low levels of MUC1*. Clinical trial information: NCT04020575 .

Updates on the mechanisms of toxicities associated with monoclonal antibodies targeting growth factor signaling and immune cells in cancer.

Monoclonal antibody (mAb)-based immunotherapy currently is considered to be an optimal therapeutic approach to cancer treatment, either in combination with surgery, radiation, and/or chemotherapy or alone. Various solid tumors and hematological malignancies have been characterized by distinct molecular targets, which could be utilized as innovative anticancer agents. Notably, receptor tyrosine kinases, including HER2, EGFR, VEGFR, and PDGFR, which act as receptors for growth factors, serve as crucial target proteins, expanding their role in the cancer therapeutic market. In contrast to conventional anticancer agents that directly target cancer cells, the advent of immunotherapy introduces novel approaches, such as immune checkpoint blockers (ICBs) and mAbs targeting surface antigens on immune cells in hematological malignancies and lymphomas. While these immunotherapies have mitigated the acquired resistance observed in traditional targeted therapies, they also exhibit diverse toxicities. Herein, this review focuses on describing the well-established toxicities and newly proposed mechanisms of monoclonal antibody toxicity in recent studies. Understanding these molecular mechanisms is indispensable to overcoming the limitations of mAbs-based therapies, facilitating the development of innovative anticancer agents, and uncovering novel indications for cancer treatment in the future.

Abstract LB307: The role of tumor microenvironment derived growth factors in pediatric brain tumors

Abstract Introduction: Children with H3K27-altered diffuse midline gliomas (DMG-H3K27a), have a 5-year survival rate of only 2% following diagnosis. While in-depth genomic characterization has been performed in DMG-H3K27a, very few druggable targets have been identified. A common feature of DMG-H3K27a is infiltration of microglia, macrophages, other myeloid cells, collectively referred to as GAMs. Cellular crosstalk between non-tumor and tumor cells in the tumor microenvironment (TME) can both promote and or inhibit tumor growth, thus representing an opportunity in the pursuit of novel therapeutics. We have recently determined that DMG-H3K27a tumor cells stimulate microglial secretion of pro-tumor growth factors. Hypothesis: Microglial-derived growth factors activate receptor tyrosine kinases (RTKs) via paracrine signaling in DMG-H3K27a. Methods: We used scRNA-seq and RNA-seq datasets and, in parallel, in vitro studies, such as co-culture studies, ELISA assays, western blotting, and growth assays to test our hypothesis. In addition, we are developing a mouse-human chimeric neutralizing antibodies of our top targets for in vivo studies. Preliminary Results: Microglial cells secrete growth factors into the TME, which promotes DMG-H3K27a cell proliferation and growth. Targeting growth factors with neutralizing antibodies attenuates glioma cell proliferation in co-culture studies. Conclusion: This research uncovers an important signaling network between GAMs and DMG-H3K27a that can be therapeutically targeted. Citation Format: Andrea Cruz, Brenden Johnson, Taylor Gatesman, Mathew Halbert, Suchet Taori, Baoli Hu, Gary Kohanbash, Ian F. Pollack, Sameer Agnihotri. The role of tumor microenvironment derived growth factors in pediatric brain tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB307.

Therapeutic Approaches for Urologic Chronic Pelvic Pain Syndrome; Management: Research Advances, Experimental Targets, and Future Directions.

Urologic chronic pelvic pain syndrome (UCPPS) is a painful chronic condition with persistent pain originating from the pelvis that often leads to detrimental lifestyle changes in the affected patients. The syndrome develops in both sexes, with an estimated prevalence of 5.7% to 26.6% worldwide. This narrative review summarizes currently recommended therapies for UCPPS, followed by the latest animal model findings and clinical research advances in the field. The diagnosis of UCPPS by clinicians has room for improvement despite the changes in the past decade aiming to decrease the time to treatment. Therapeutic approaches targeting growth factors (i.e., nerve growth factor, vascular endothelial growth factor), amniotic bladder therapy, and stem cell treatments gain more attention as experimental treatment options for UCPPS. The development of novel diagnostic tests based on the latest advances in urinary biomarkers would be beneficial to assist with the clinical diagnosis of UCPPS. Future research directions should address the role of chronic psychologic stress and the mechanisms of pain refractory to conventional management strategies in UCPPS etiology. Testing the applicability of cognitive behavioral therapy in this cohort of UCPPS patients might be promising to increase their quality of life. The search for novel lead compounds and innovative drug delivery systems requires clinically relevant translational animal models. The role of autoimmune responses triggered by environmental factors is another promising research direction to clarify the impact of the immune system in UCPPS pathophysiology. SIGNIFICANCE STATEMENT: This minireview provides an up-to-date summary of the therapeutic approaches for UCPPS with a focus on recent advancements in the clinical diagnosis and treatments of the disease, pathophysiological mechanisms of UCPPS, signaling pathways, and molecular targets involved in pelvic nociception.

Novel bisphosphonate-based cathepsin K-triggered compound targets the enthesis without impairing soft tissue-to-bone healing.

Background: Osteoadsorptive fluorogenic sentinel 3 (OFS-3) is a recently described compound that contains a bone-targeting bisphosphonate (BP) and cathepsin K (Ctsk)-triggered fluorescence signal. A prior study in a murine Achilles repair model demonstrated its effectiveness at targeting the site of tendon-to-bone repair, but the intrinsic effect of this novel bisphosphonate chaperone on tendon-to-bone healing has not been previously explored. We hypothesized that application of this bisphosphonate-fluorophore cargo conjugate would not affect the biomechanical properties or histologic appearance of tendon-bone repairs. Materials and Methods: Right hindlimb Achilles tendon-to-bone repair was performed on 12-week old male mice. Animals were divided into 2 groups of 18 each: 1) Achilles repair with OFS-3 applied directly to the repair site prior to closure, and 2) Achilles repair with saline applied prior to closure. Repaired hindlimbs from 12 animals per group were harvested at 6 weeks for biomechanical analysis with a custom 3D-printed jig. At 4 and 6 weeks, repaired hindlimbs from the remaining animals were assessed histologically using H&E, immunohistochemistry (IHC) staining for the presence of Ctsk, and second harmonic generation (SHG) imaging to evaluate collagen fibers. Results: At 6 weeks, there was no significant difference in failure load, stiffness, toughness, or displacement to failure between repaired hindlimbs that received OFS-3 versus saline. There was no difference in tissue healing on H&E or Ctsk staining on immunohistochemistry between animals that received OFS-3 versus saline. Finally, second harmonic generation imaging demonstrated no difference in collagen fiber parameters between the two groups. Conclusion: OFS-3 did not significantly affect the biomechanical properties or histologic appearance of murine Achilles tendon-to-bone repairs. This study demonstrates that OFS-3 can target the site of tendon-to-bone repair without causing intrinsic negative effects on healing. Further development of this drug delivery platform to target growth factors to the site of tendon-bone repair is warranted.

Golgi Protein 73 Promotes Angiogenesis in Hepatocellular Carcinoma.

Golgi protein 73 (GP73), a resident protein of the Golgi apparatus, is notably elevated in hepatocellular carcinoma (HCC). While its critical role in remodeling the tumor microenvironment (TME) is recognized, the intricate mechanisms are not fully understood. This study reveals that GP73 in HCC cells interacts with prolyl hydroxylase-2 (PHD-2) in a competitive manner, thereby impeding the hydroxylation of hypoxia-induced factor-1α (HIF-1α). The effect above promotes the production and secretion of vascular endothelial growth factor A (VEGFA). Moreover, exosomal GP73 derived from HCC cells can be internalized by human umbilical vein endothelial cells (HUVECs) and competitively interact with HECTD1, an E3 ubiquitin ligase targeting growth factor receptor-bound protein 2 (GRB2). This interaction stabilizes GRB2, thereby activating the Ras-mitogen-activated protein kinase (MAPK) signaling pathway. Consequently, escalated levels of GP73 intensify VEGF production in HCC cells and potentiate mitogenic signaling in vascular endothelial cells, fostering angiogenesis in the TME. Our findings propose that GP73 might serve as a novel target for anti-angiogenic therapy in HCC.

Pigmented Microbial Extract (PMB) from Exiguobacterium Species MB2 Strain (PMB1) and Bacillus subtilis Strain MB1 (PMB2) Inhibited Breast Cancer Cells Growth In Vivo and In Vitro.

Breast cancer (BC) continues to be one of the major causes of cancer deaths in women. Progress has been made in targeting hormone and growth factor receptor-positive BCs with clinical efficacy and success. However, little progress has been made to develop a clinically viable treatment for the triple-negative BC cases (TNBCs). The current study aims to identify potent agents that can target TNBCs. Extracts from microbial sources have been reported to contain pharmacological agents that can selectively inhibit cancer cell growth. We have screened and identified pigmented microbial extracts (PMBs) that can inhibit BC cell proliferation by targeting legumain (LGMN). LGMN is an oncogenic protein expressed not only in malignant cells but also in tumor microenvironment cells, including tumor-associated macrophages. An LGMN inhibition assay was performed, and microbial extracts were evaluated for in vitro anticancer activity in BC cell lines, angiogenesis assay with chick chorioallantoic membrane (CAM), and tumor xenograft models in Swiss albino mice. We have identified that PMB from the Exiguobacterium (PMB1), inhibits BC growth more potently than PMB2, from the Bacillus subtilis strain. The analysis of PMB1 by GC-MS showed the presence of a variety of fatty acids and fatty-acid derivatives, small molecule phenolics, and aldehydes. PMB1 inhibited the activity of oncogenic legumain in BC cells and induced cell cycle arrest and apoptosis. PMB1 reduced the angiogenesis and inhibited BC cell migration. In mice, intraperitoneal administration of PMB1 retarded the growth of xenografted Ehrlich ascites mammary tumors and mitigated the proliferation of tumor cells in the peritoneal cavity in vivo. In summary, our findings demonstrate the high antitumor potential of PMB1.

Growth factors and female reproduction in vertebrates

Female reproductive efficiency is influenced by the outcomes of various processes, including folliculogenesis, apoptosis, response to gonadotropin signaling, oocyte maturation, and ovulation. The role of hormones in regulating these processes and other reproductive activities has been well established. It is becoming increasingly evident that in addition to well-characterized hormones, growth factors play vital roles in regulating some of these reproductive activities. Growth factors and their receptors are widely distributed in vertebrate ovaries at different stages of ovarian development, indicating their involvement in intraovarian reproductive functions. In the ovary, cell surface receptors allow growth factors to regulate intraovarian reproductive activities. Understanding these actions in the reproductive axis would provide a tool to target growth factors and/or their receptors to yield desirable reproductive outcomes. These include enrichment of in vitro maturation and fertilization culture media, and management of infertility. This review discusses some widely characterized growth factors belonging to the TGF, EGF, IGF, FGF, and BDNF family of peptides and their role in female reproduction in vertebrates, with a focus on mammals.

MOLECULAR DOCKING AND INVESTIGATION OF BOSWELLIA SERRATA PHYTOCOMPOUNDS AS CANCER THERAPEUTICS TO TARGET GROWTH FACTOR RECEPTORS: AN IN SILICO APPROACH

Objective: Boswellia serrata is a plant with a long history of use in traditional medicine, particularly for its anti-inflammatory and anti-cancer properties. Growth factors and their receptors are significant components in the initiation and progression of malignancy, and aberrant functioning of these pathways can result in unrestrained cell division and expansion. Methods: In this study, an in silico approach was used to explore the potential of Boswellia serrata phytochemicals as cancer therapeutics to target growth factor receptors. The virtual screening involved molecular docking simulations (PyRx) to predict the binding affinity between the phytochemicals and the receptors. Results: The seventy-four phytocompounds identified from Boswellia serrata were preliminarily screened based on their binding towards growth factor receptors. The ligands demonstrated better binding with the GFR targets, and the binding score less than-7 kcal/mol was considered for further investigation results demonstrated that Alpha-boswellic exhibited strong binding affinity to the receptors, suggesting their potential as targeted cancer therapies. This study provides a foundation for future in vitro and in vivo experiments to validate the efficacy of these phytochemicals as cancer treatments. Conclusion: The results suggest that Boswellic acid derivatives from Boswellia serrata could be a promising source of new cancer therapies.

Optimizing Delivery of Therapeutic Growth Factors for Bone and Cartilage Regeneration

Bone- and cartilage-related diseases, such as osteoporosis and osteoarthritis, affect millions of people worldwide, impairing their quality of life and increasing mortality. Osteoporosis significantly increases the bone fracture risk of the spine, hip, and wrist. For successful fracture treatment and to facilitate proper healing in the most complicated cases, one of the most promising methods is to deliver a therapeutic protein to accelerate bone regeneration. Similarly, in the setting of osteoarthritis, where degraded cartilage does not regenerate, therapeutic proteins hold great promise to promote new cartilage formation. For both osteoporosis and osteoarthritis treatments, targeted delivery of therapeutic growth factors, with the aid of hydrogels, to bone and cartilage is a key to advance the field of regenerative medicine. In this review article, we propose five important aspects of therapeutic growth factor delivery for bone and cartilage regeneration: (1) protection of protein growth factors from physical and enzymatic degradation, (2) targeted growth factor delivery, (3) controlling GF release kinetics, (4) long-term stability of regenerated tissues, and (5) osteoimmunomodulatory effects of therapeutic growth factors and carriers/scaffolds.

Comprehensive assessment of growth factors, inflammatory mediators, and cytokines in vitreous from patients with proliferative diabetic retinopathy

To assess alterations in growth factors, inflammatory mediators, and cytokines associated with vitreous-retinal diseases in vitreous humor from patients with proliferative diabetic retinopathy (PDR), and to identify potential new treatment targets and strategies. Control vitreous samples were collected from patients with macular hole, epiretinal membranes, or rhegmatogenous retinal detachments, and PDR samples from patients with complications of PDR, who required pars plana vitrectomy. Specimens were stored at -80°C and then investigated by Luminex multi-factor assay. Parametric and nonparametric analyses of demographic characteristics and cytokine expression levels were conducted using SPSS. There were no significant differences in demographic characteristics between patients with and without PDR. Expression levels of growth factors [platelet-derived growth factor (PDGF)-AA, glial cell line-derived neurotrophic factor (GDNF), and vascular endothelial growth factor A (VEGFA)], inflammatory mediators [interleukin (IL)-8, IL-11, and tumor necrosis factor-α (TNF-α)] and cytokines [chemokine C-X-C ligand (CXCL)10, interferon-γ (IFN-γ), and granulocyte macrophage-colony stimulating factor (GM-CSF)] were significantly elevated in vitreous humor from patients with PDR compared with those in the control group (all P<0.05). Further, VEGFA levels were lower in patients with PDR treated with anti-VEGF injection than those who were not (P<0.05), and there was no difference between the PDR group treated with anti-VEGF and controls (P>0.05). This proof-of-concept study demonstrates the potential for combinational therapeutic strategies to ameliorate diabetic retinopathy progression by targeting growth factors, inflammatory factors, and cytokines, in addition to VEGFA.

Non-Destructive Monitoring of Crop Fresh Weight and Leaf Area with a Simple Formula and a Convolutional Neural Network.

Crop fresh weight and leaf area are considered non-destructive growth factors due to their direct relation to vegetative growth and carbon assimilation. Several methods to measure these parameters have been introduced; however, measuring these parameters using the existing methods can be difficult. Therefore, a non-destructive measurement method with high versatility is essential. The objective of this study was to establish a non-destructive monitoring system for estimating the fresh weight and leaf area of trellised crops. The data were collected from a greenhouse with sweet peppers (Capsicum annuum var. annuum); the target growth factors were the crop fresh weight and leaf area. The crop fresh weight was estimated based on the total system weight and volumetric water content using a simple formula. The leaf area was estimated using top-view images of the crops and a convolutional neural network (ConvNet). The estimated crop fresh weight and leaf area exhibited average R2 values of 0.70 and 0.95, respectively. The simple calculation was able to avoid overfitting with fewer limitations compared with the previous study. ConvNet was able to analyze raw images and evaluate the leaf area without additional sensors and features. As the simple calculation and ConvNet could adequately estimate the target growth factors, the monitoring system can be used for data collection in practice owing to its versatility. Therefore, the proposed monitoring system can be widely applied for diverse data analyses.

MicroRNA-223 negatively regulates the osteogenic differentiation of periodontal ligament derived cells by directly targeting growth factor receptors

BackgroundMicroRNA (miRNA) is accepted as a critical regulator of cell differentiation. However, whether microRNA-223 (miR-223) could affect the osteogenic differentiation of periodontal ligament (PDL)-derived cells is still unknown. The aim of this study was to explore the mechanisms underlying the roles of miR-223 in the osteogenesis of PDL-derived cells in periodontitis.MethodsMicroarray analysis and real-time polymerase chain reaction (RT-PCR) were used to identify difference in miR-223 expression pattern between healthy and inflamed gingival tissue. The target genes of miR-223 were predicted based on Targetscan and selected for enrichment analyses based on Metascape database. The gain-and loss-of-function experiments were performed to discuss roles of miR-223 and growth factor receptor genes in osteogenic differentiation of PDL-derived cells. The target relationship between miR-223 and growth factor receptor genes was confirmed by a dual luciferase assay. Osteogenic differentiation of PDL-derived cells was assessed by Alizarin red staining, RT-PCR and western blot detection of osteogenic markers, including osteocalcin (OCN), osteopontin (OPN) and runt-related transcription factor 2 (Runx2).ResultsMiR-223 was significantly increased in inflamed gingival tissues and down-regulated in PDL-derived cells during osteogenesis. The expression of miR-223 in gingival tissues was positively correlated with the clinical parameters in periodontitis patients. Overexpression of miR-223 markedly inhibited PDL-derived cells osteogenesis, which was evidenced by reduced Alizarin red staining and osteogenic markers expressions. Furthermore, two growth factor receptor genes, including fibroblast growth factor receptor 2 (FGFR2) and transforming growth factor beta receptor 2 (TGFβR2), were revealed to be direct targets of miR-223 and shown to undergo up-regulation in PDL-derived cells during osteogenesis. Moreover, suppression of FGFR2 or TGFβR2 dramatically blocked PDL-derived cells osteogenic differentiation.ConclusionsOur study provides novel evidence that miR-223 can be induced by periodontitis and acts as a negative regulator of PDL-derived cells osteogenesis by targeting two growth factor receptors (TGFβR2 and FGFR2).

Abstract P3070: Distinct Inflammatory Milieu In Patients With Right Heart Failure

Introduction: Right heart failure (RHF) is associated with worse clinical outcomes in patients with heart failure (HF) and pulmonary hypertension. Previous studies have shown elevated circulating proinflammatory cytokines in patients with HF; however, no study has compared the inflammatory signature in patients with RHF to those with HF and preserved right ventricular (RV) function. Hypothesis: We predicted that patients with RHF would have a distinct inflammatory milieu compared to normal and HF patients. Methods: We recruited adult patients undergoing right heart catheterization (RHC) and excluded those with a history of transplant, autoimmune disease, malignancy, or infection. Patients underwent blood sampling during RHC and were classified into normal, HF, and RHF cohorts a priori . We then performed multiplex protein assay on serum targeting cytokines, chemokines, and growth factors. Results: A total of 43 patients were included. HF and RHF patients had similar demographics and cardiac indices, but the RHF cohort had elevated markers of congestion characterized by higher right atrial pressure and creatinine, and a trend towards higher bilirubin (Figure). Patients with RHF had elevated levels of select proinflammatory cytokines, chemokines, and growth factors. Of the factors measured, VEGF-A, soluble CD163, and CXCL12 were predictive of all-cause mortality and LVAD/transplant-free survival. Conclusion: Patients with RHF have a distinct proinflammatory signature compared to those with preserved RV function. VEGF-A, soluble CD163, and CXCL12 predicted clinical outcomes in an advanced HF population.

Targeting Growth Factor and Cytokine Pathways to Treat Idiopathic Pulmonary Fibrosis.

Idiopathic pulmonary fibrosis (IPF) is a chronic interstitial lung disease of unknown origin that usually results in death from secondary respiratory failure within 2–5 years of diagnosis. Recent studies have identified key roles of cytokine and growth factor pathways in the pathogenesis of IPF. Although there have been numerous clinical trials of drugs investigating their efficacy in the treatment of IPF, only Pirfenidone and Nintedanib have been approved by the FDA. However, they have some major limitations, such as insufficient efficacy, undesired side effects and poor pharmacokinetic properties. To give more insights into the discovery of potential targets for the treatment of IPF, this review provides an overview of cytokines, growth factors and their signaling pathways in IPF, which have important implications for fully exploiting the therapeutic potential of targeting cytokine and growth factor pathways. Advances in the field of cytokine and growth factor pathways will help slow disease progression, prolong life, and improve the quality of life for IPF patients in the future.

MiR-320 regulates myogenesis by targeting growth factor receptor-bound protein-2 and ameliorates myotubes atrophy.

Loss of muscle mass can lead to diseases such as sarcopenia, diabetes, and obesity, which can worsen the quality of life and increase the incidence of disease. Therefore, understanding the mechanism underlying skeletal muscle differentiation is vital to prevent muscle diseases. We previously found that microRNA-320 (miR-320) is highly expressed in the lean muscle-type pigs, but its regulatory role in myogenesis remains unclear. The bioinformatics prediction indicated that miR-320 could bind to the 3 'untranslated region of growth factor receptor-bound protein-2 (Grb2). We hypothesized that miR-320 targets Grb2 to regulate myoblasts differentiation. To verify this, we transfected miR-320 mimic and inhibitor into C2C12 myoblasts to assess the role of miR-320 during myoblasts differentiation. We used real-time qPCR, luciferase reporter assays, and western blotting to confirm that miR-320 directly targets Grb2 to promote myoblasts differentiation. Moreover, by using a dexamethasone-induced atrophic model of myotubes, we discovered that miR-320 promotes the repair of damaged myotubes. Our findings expand understanding of miRNAs and genes related to regulating skeletal muscle differentiation, and provide insight into underlying therapeutic strategies for muscle diseases.

New approaches in ovarian cancer based on genetics and carcinogenesis hypotheses (Review).

Ovarian cancer is the leading cause of death among gynecological malignancies and its incidence is rising in the last decades especially in developed countries. High-grade serous ovarian cancer (HGSOC) represents 70% of ovarian cancers. Oral contraceptive use and salpingo-oophorectomy or salpingectomy are well known protective factors against development of ovarian cancer. Identification of specific mutations associated with a high risk of developing ovarian cancer, especially BRCA1/2 mutation and TP53 mutations, has paved the way for implementation of new strategies for early diagnosis and therapy. Hereditary forms of ovarian cancer account for 5-10% and have BRCA1/2 gene mutations or TP53 mutations. BRCA1/2 gene mutations appear in 22% of HGSOC and are associated with the defective homologous repair (HR)/DNA repair pathway. Genetic testing in ovarian cancer is important for risk assessment and therapeutic options. Although 'universal genetic testing' is not recommended yet, the procedure remains highly recommended in women with high risk. Genes involved in the development of ovarian cancer as TP53 may be targeted by gene therapy. Poly (ADP-ribose) polymerase (PARP) inhibitors may enhance the cytotoxic effect of DNA-damaging chemotherapy, and induce synthetic lethality in cases with BRCA1/2 mutations. Other strategies are designed to target pathways driven by various gene mutations, including the use of tyrosine kinase inhibitors in low-grade serous ovarian cancer (LGSOC), or the use of drugs, which target growth factors, or epigenetic events including methylation, and acetylation of genes. The tubal involvement in ovarian carcinogenesis provides an important tool for the clinician to implement risk-reducing strategies including salpingo-oophorectomy or salpingectomy in high-risk cases at appropriate ages.