Plasma TGF-β1 Research Articles

Evidence for platelet-derived transforming growth factor β1 as an early inducer of liver regeneration after hepatectomy in mice.

Platelets play a crucial role in tissue regeneration, and their involvement in liver regeneration is well-established. However, the specific contribution of platelet-derived Transforming Growth Factor Beta 1 (TGFβ1) to liver regeneration remains unexplored. This study investigated the role of platelet-derived TGFβ1 in initiating liver regeneration following 2/3 liver resection. Using platelet-specific TGFβ1 knockout (Plt.TGFβ1 KO) mice and wild-type littermates (Plt.TGFβ1 WT) as controls, the study assessed circulating levels and hepatic gene expression of TGFβ1, Platelet Factor 4 (PF4), and Thrombopoietin (TPO) at early time points post-hepatectomy (post-PHx). Hepatocyte proliferation was quantified through Ki67 staining and PCNA expression in total liver lysates at various intervals, and phosphohistone-H3 (PHH3) staining was employed to mark mitotic cells. Circulating levels of hepatic mitogens, Hepatocyte Growth Factor (HGF), and Interleukin-6 (IL6) were also assessed. Results revealed that platelet-TGFβ1 deficiency significantly reduced total plasma TGFβ1 levels at 5 h post-PHx in Plt.TGFβ1 KO mice compared to controls. While circulating PF4 levels, liver platelet recruitment and activation appeared normal at early time points, Plt.TGFβ1 KO mice showed more stable circulating platelet numbers with higher numbers at 48 h post-PHx. Notably, hepatocyte proliferation was significantly reduced in Plt.TGFβ1 KO mice. The results show that a lack of TGFβ1 in platelets leads to an unbalanced expression of IL6 in the liver and to strongly increased HGF levels 48 h after liver resection, and yet liver regeneration remains reduced. The study identifies platelet-TGFβ1 as a regulator of hepatocyte proliferation and platelet homeostasis in the early stages of liver regeneration.

Immunoregulatory Cells and Cytokines Discriminate Disease Activity Score 28-Remission Statuses and Ultrasound Grades in Rheumatoid Arthritis Patients with Non-High Disease Activity.

It is not clear whether immunoregulatory cytokines and cells are associated with Disease Activity Score 28 (DAS28) scores and ultrasound grades/scores. Here, we investigated the relationships between immunoregulatory cytokines or cells and different DAS28 scores or ultrasound grades/scores in patients with rheumatoid arthritis (RA). This study enrolled 50 RA patients (with 147 visits) who had remission/low/moderate DAS28-ESR scores (92% in remission and low disease activity) at baseline. Blood was collected and an ultrasound was performed three times in a year. Percentages of regulatory B cells and T regulatory type 1 cells and M2 macrophage numbers in the blood were examined. Plasma levels of 10 immunoregulatory cytokines IL-4, IL-5, IL-9, IL-10, IL-13, IL-27, IL-35, TGF-β1, sTNF-R1, and sTNF-R2 and monocyte chemotactic protein-1 (MCP-1) were assessed using ELISA assay. The correlations of cytokines and cells with different DAS28 scores and ultrasound grades were investigated, and cytokines and cells were compared between different categories of DAS28 scores and ultrasound grades. Plasma TGF-β1 levels were higher in the DAS28-ESR < 2.6 (remission) subgroup than in the DAS28-ESR ≥ 2.6 (nonremission) subgroup (p = 0.037). However, plasma TGF-β1 levels were higher in the high ultrasound grade subgroup than those in the low ultrasound grade subgroup (p = 0.007). The number of M2 macrophages was lower in the DAS28-MCP-1 < 2.2 subgroup than in the DAS28-MCP-1 ≥ 2.2 subgroup (p = 0.036). The levels of TGF-β1, sTNF-R2, IL-10, and IL-27 were higher in patients with high ultrasound grades than in those with low ultrasound grades. IL-27 was also higher in the nonremission DAS28-ESR subgroup than the remission one (p = 0.025). Moreover, sTNF-R1 levels in the 2011 American College of Rheumatology/European League Against Rheumatism (ACR/EULAR) remission subgroup were significantly lower than in the 2011 ACR/EULAR nonremission subgroup (p = 0.007). This trend was reflected in that lower sTNF-R1 levels correlated with low DAS28-MCP-1 scores (rho = 0.222, p = 0.007). We conclude that high plasma TGF-β1 levels indicate the DAS28-ESR remission (<2.6) subgroup and the high ultrasound grade subgroup. IL-27 probably connects the nonremission DAS28-ESR to high ultrasound grades. Low sTNF-R1 levels probably link low DAS28-MCP-1 scores with the 2011 ACR/EULAR remission subgroup. It suggests that incongruent immuno-inflammatory abnormalities exist between DAS28 scores and ultrasound grades, and are also dissimilar among various DAS28-formula categories. Therefore, this study may provide a basis for further research into individual cytokines and immunoregulatory cells behind each DAS28 formula and ultrasound grades/scores.

Burkholderia cepacia in cystic fibrosis children and adolescents: overall survival and immune alterations.

In current literature there are only scarce data on the host inflammatory response during Burkholderia cepacia complex (Bcc) persistence. The primary objective of the present research was to carry out cross-sectional analyses of biomarkers and evaluate disease progression in cystic fibrosis (CF) patients with chronic Bcc infection and pathogen-free ones. The secondary aim was to assess prospectively overall survival of the study participants during up to 8 years of follow-up. The study included 116 paediatric patients with CF; 47 CF patients were chronically infected with Bcc, and 69 individuals were Bcc free. Plasma and sputum biomarkers (neutrophil elastase, MMP-8, MMP-9, MMP-12, IL-2, IL-4, IL-6, IL-8, IL-10, IL-18, IL-22, IL-23, IL-17, IFN-γ, TGFβ1, TNF-α) were analysed using commercially available kits. Besides, inhibitory effect of dexamethasone on proliferative response of PHA-stimulated peripheral blood lymphocytes had been assessed. Bcc infected patients did not differ from Bcc free ones in demographic and clinical parameters, but demonstrated an increased rate of glucose metabolism disturbances and survival disadvantage during prolong follow-up period. Biomarkers analyses revealed elevated TNF-α and reduced IL-17F levels in sputum samples of Bcc infected patients. These patients also demonstrated improvement of peripheral blood lymphocyte sensitivity to steroid treatment and reduction in plasma pro-inflammatory (IL-17F and IL-18) and anti-inflammatory (TGFβ1 and IL-10) cytokine concentrations. Reduction in IL-17F levels may have several important consequences including increase in steroid sensitivity and glycemic control disturbances. Further investigations are needed to clarify the role of IL-17 cytokines in CF complication development. Low plasma TGFβ1 and IL-10 levels in Bcc infected group may be a sign of subverted activity of regulatory T cells. Such immune alterations may be one of the factors contributing to the development of the cepacia syndrome.

ILC2s induce Treg but not Th2-type immunity through IL-33/ST2 pathway in pulmonary tuberculosis.

We investigated the function of type 2 innate lymphoid cells (ILC2s) and IL-33 in pulmonary tuberculosis (PTB). Peripheral blood samples were collected from PTB patients and healthy controls. The cytometric bead array was used to detect plasma IL-33, TGF-β, IL-4, IL-5, IL-6, IL-10, IL-13, and soluble ST2 (sST2). ILC2s, Th2, and Treg cells were detected with flow cytometry. Quantitative real-time PCR was used to measure mRNA levels. ILC2s were co-cultured with peripheral blood mononuclear cells and then intervened with IL-33 or anti-ST2 antibody + IL-33 in vitro. IL-4, IL-6, IL-5, IL-10, IL-13, and TGF-β levels were measured by enzyme-linked immunosorbent assay. Compared with healthy controls, the levels of IL-33, sST2, TGF-β, IL-10, and IL-6 in the plasma of PTB patients were significantly higher. No significant difference was found in the plasma IL-4, IL-5, and IL-13 levels. Patients with PTB had significantly increased ILC2s proportion and mRNA levels of RAR-related orphan receptor α and GATA binding protein 3. After 48 h of IL-33 stimulation in vitro, Treg cell proportion significantly increased and the IL-10 level was significantly elevated. Treatment with anti-ST2 abolished these effects. No significant difference was found in cytokines of IL-4, IL-6, IL-5, IL-13, and TGF-β, or Th2 cells before and after IL-33 treatment. ILC2s proportion in peripheral blood was increased and plasma IL-33 was upregulated in PTB patients. IL-33 may promote the growth of ILC2s and the production of Treg-related cell cytokines, but not Th2-related cell cytokines, to participate in immune response to PTB.

A pilot study on the mechanism of Guasha in treating Parkinson's disease based on molecular level and ultra-trace proteomics analysis

Guasha is a widely applied non-pharmacological therapy of traditional Chinese medicine (TCM), which has been proved to be effective in the treatment of Parkinson's disease (PD). The objective of this study was to explore the mechanisms of Guasha therapy in the pathological processes including neuroinflammation in PD. After 3 Guasha courses, a reduction in α-synuclein aggregation and alleviation of microglia activation were observed in the substantia nigra pars compacta (SNpc). Furthermore, levels of TNF-α, IL-1β and IFN-γ in the SNpc decreased, while plasma TGF-β, IL-4 and IL-10 increased. Moreover, both TH protein and mRNA levels in the SNpc, as well as dopamine levels in the striatum, exhibited an increase. The proteomics analysis results based on plasma-derived exosomes revealed a total of 943 differentially expressed proteins identified. Compared to the model group, the Guasha group screened for 82 differential proteins, with 30 upregulated and 52 downregulated. The improvement of pathological changes in the PD model mice treated with Guasha primarily involves biological processes such as oxidative stress, immune response and inflammation, and cellular structure regulation. It involves signaling pathways related to aldosterone synthesis and secretion, adrenergic signaling in myocardial cells, COVID-19-related inflammatory signaling, and neurotrophic signaling. Collectively, the mechanisms of Guasha for treating PD might be closely related to inhibiting microglial cell activation-mediated neuroinflammation, regulating oxidative stress, cellular structure, aldosterone synthesis and secretion-mediated electrolyte balance, as well as noradrenergic signaling-mediated neuroprotection. These findings provided new insight for Guasha in treating PD and would potentially enhance therapeutic interventions.

Investigating the role of TGF-β and BDNF in cancer-related depression: a primary cross-sectional study.

Cancer-related depression is a well-documented condition that significantly impacts long-term quality of life. Brain-derived neurotrophic factor (BDNF), a neurotrophin essential for neurogenesis and neuronal plasticity, has been implicated in various neuropsychological disorders including depression associated with cancer. Cytokines, on the other hand, play a crucial role in regulating depression, potentially by influencing BDNF expression. Transforming growth factor-β (TGF-β), a key immune regulator within the tumor microenvironment, has been found to elevate BDNF levels, establishing a link between peripheral immune responses and depression. The study aims to investigate the correlation of TGF-β and BDNF in cancer-related depression. This study involved a cohort of 153 gynecological patients, including 61 patients with gynecological cancer and 92 patients without cancer. Depression levels were assessed using the subscale of Hospital Anxiety and Depression Scale (HADS-D), and TGF-β and BDNF plasma levels were measured using enzyme-linked immunosorbent assay (ELISA). The study revealed elevated plasma TGF-β levels in patients with cancer (32.24 ± 22.93 ng/ml) compared to those without cancer (25.24 ± 19.72 ng/ml) (P = 0.046). Additionally, reduced levels of BDNF were observed in patients presenting depression symptoms (44.96 ± 41.06 pg/ml) compared to those without depression (133.5 ± 176.7 pg/ml) (P = 0.036). Importantly, a significant correlation between TGF-β and BDNF was found in patients without cancer but with depression (correlation coefficient = 0.893, **P < 0.01). Interestingly, cancer appeared to influence the association between TGF-β and BDNF in patients with depression, as evidenced by a significant difference in the correlation of TGF-β and BDNF between cancer and non-cancer groups (P = 0.041). These findings underscore the active involvement of TGF-β and BDNF crosstalk in the context of cancer-related depression.

Low TGF-β1 plasma levels are associated with cognitive decline in Down syndrome.

Almost all individuals with Down's syndrome (DS) show the characteristic neuropathological features of Alzheimer's disease (AD) by the age of 40, yet not every individual with DS experiences symptoms of AD later in life. Similar to neurotypical developing subjects, AD in people with DS lasts for a long preclinical phase in which biomarkers follow a predictable order of changes. Hence, a prolonged asymptomatic period precedes the onset of dementia, underscoring the importance of identifying new biomarkers for the early detection and monitoring of cognitive decline in individuals with DS. Blood-based biomarkers may offer an alternative non-invasive strategy for the detection of peripheral biological alterations paralleling nervous system pathology in an early phase of the AD continuum. In the last few years, a strong neurobiological link has been demonstrated between the deficit of transforming growth factor-β1 (TGF-β1) levels, an anti-inflammatory cytokine endowed with neuroprotective activity, and early pro-inflammatory processes in the AD brain. In this clinical prospective observational study, we found significant lower plasma TGF-β1 concentrations at the first neuropsychological evaluation (baseline = T0) both in young adult DS individuals (19-35years) and older DS subjects without AD (35-60years) compared to age- and sex-matched healthy controls. Interestingly, we found that the lower TGF-β1 plasma concentrations at T0 were strongly correlated with the following cognitive decline at 12 months. In addition, in young individuals with DS, we found, for the first time, a negative correlation between low TGF-β1 concentrations and high TNF-α plasma concentrations, a pro-inflammatory cytokine that is known to be associated with cognitive impairment in DS individuals with AD. Finally, adopting an ex vivo approach, we found that TGF-β1 concentrations were reduced in parallel both in the plasma and in the peripheral blood mononuclear cells (PBMCs) of DS subjects, and interestingly, therapeutic concentrations of fluoxetine (FLX) applied to cultured PBMCs (1µM for 24h) were able to rescue TGF-β1 concentrations in the culture media from DS PBMCs, suggesting that FLX, a selective serotonin reuptake inhibitor (SSRI) endowed with neuroprotective activity, might rescue TGF-β1 concentrations in DS subjects at higher risk to develop cognitive decline.

LINC00158 modulates the function of BEAS-2B cells via targeting BCL11B and ameliorates OVA-LPS-induced severe asthma in mice models

Persistent type (T) 2 airway inflammation plays an important role in the development of severe asthma. However, the molecular mechanisms leading to T2 severe asthma have yet to be fully clarified. Human normal lung epithelial cells (BEAS-2B cells) were transfected with LINC00158/BCL11B plasmid/small interfering RNA (siRNA). Levels of epithelial-mesenchymal transition (EMT)-related markers were measured using real-time qPCR (RT-qPCR) and western blot. A dual luciferase reporter assay was used to validate the targeting relationship between LINC00158 and BCL11B. The effects of LINC00158-lentivirus vector-mediated overexpression and dexamethasone on ovalbumin (OVA)/lipopolysaccharide (LPS)-induced severe asthma were investigated in mice in vivo. Our study showed that overexpression of LINC00158/BCL11B inhibited the levels of EMT-related proteins, apoptosis, and promoted the proliferation of BEAS-2B cells. BCL11B was a direct target of LINC00158. And LINC00158 targeted BCL11B to regulate EMT, apoptosis, and cell proliferation of BEAS-2B cells. Compared with severe asthma mice, LINC00158 overexpression alleviated OVA/LPS-induced airway hyperresponsiveness and airway inflammation, including reductions in T helper 2 cells factors in lung tissue and BALF, serum total- and OVA-specific IgE, inflammatory cell infiltration, and goblet cells hyperplasia. In addition, LINC00158 overexpression alleviated airway remodeling, including reduced plasma TGF-β1 and collagen fiber deposition, as well as suppression of EMT. Additionally, overexpression of LINC00158 enhanced the therapeutic effect of dexamethasone in severe asthmatic mice models. LINC00158 regulates BEAS-2B cell biological function by targeting BCL11B. LINC00158 ameliorates T2 severe asthma in vivo and provides new insights into the clinical treatment of severe asthma.

Adipose-derived stem cell-derived exosomes regulate Th2/Treg balance in peripheral blood of AR patients through the mTOR pathway

Objective:To investigate the mechanism of adipose derived stem cell exosomes（ADSC-exos） regulating Th2/Treg balance in peripheral blood of patients with allergic rhinitis（AR）. Methods:Thirty patients with AR who were treated in Department of Otolaryngology Head and Neck Surgery, the First Affiliated Hospital of Zhengzhou University from March 2022 to October 2022 were selected, and 30 patients with simple deviation of nasal septum who were treated in our department during the same period were selected as the control group. 10 mL peripheral venous blood was collected from all patients. The levels of IL-4 and TGF-β in plasma were analyzed by ELISA. PBMCs were isolated by density gradient centrifugation. Then, protein and RNA were further extracted, and the expression levels of IL-4, TGF-β, GATA3 and Foxp3 genes were detected by qRT-PCR. Western Blotting detected p-PI3K（P85）, p-AKT（Ser473） in PBMCs of AR patients and healthy controls. Protein expression levels of p-mTOR（Ser2448）, p-p70S6K（Thr389）, and the proportion of Th2 and Treg cells were analyzed by flow cytometry. PBMCs of AR patients were stimulated to differentiate and co-cultured with exosomes of adipose stem cells. p-PI3K（P85）, p-AKT（Ser473）, p-mTOR（Ser2448） were detected in exosome treated group and untreated group by Western Blotting. The expression level of p-p70S6K（Thr389） protein, the proportion of Th2 and Treg cells were analyzed by flow cytometry, and the levels of IL-4 and TGF-β in the supernatant of cell culture were detected by ELISA. Results:Compared with the control group, the mTOR pathway in peripheral blood of AR group was significantly activated, the level of IL-4 in plasma was increased, and the level of TGF-β was decreased（P<0.05）. Compared with the control group, the proportion of Th2 cells in peripheral blood was increased, and the proportion of Treg cells was decreased（P<0.01）. Compared with the untreated group, the expression level of mTOR pathway protein decreased, the level of IL-4 decreased, and the level of TGF-β increased. The proportion of Th2 cells decreased, and the proportion of Treg cells increased（P<0.01）. Conclusion:There is an imbalance of Th2 and Treg cells in peripheral blood mononuclear cells of AR patients; the PI3K/AKT/mTOR/p70S6K pathway is activated in peripheral blood mononuclear cells of AR patients Exosomes derived from adipose mesenchymal stem cells may regulate Th2/Treg balance in AR patients through the PI3K/AKT/mTOR/p70S6K pathway.

Plasma MCP-1 and TGF-β1 Levels are Associated with Kidney Injury in Children with Congenital Anomalies of the Kidney and Urinary Tract.

Congenital anomalies of the kidney and urinary tract (CAKUT) are primarily causal for end-stage renal disease and have significant implications for long-term survival. A total of 39 healthy controls and 94 children with chronic kidney disease (CKD) were enrolled (3-12 years old as children, 13-18 years old as adolescents), who were divided into CAKUT and Non-CAKUT according to the etiology of CKD. CKD group was further classified according to estimating glomerular filtration rate (eGFR). Circulating levels of inflammatory markers such as interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), monocyte chemokine-1 (MCP-1), and transforming growth factor-β1 (TGF-β1) were analyzed. The relationship between these inflammatory markers with eGFR and the kidney injury parameter (urine protein) was investigated to assess their potential as early markers of disease progression. All circulating levels of these inflammatory cytokines were increased in CKD patients (including CAKUT and Non-CAKUT) compared with healthy subjects. The circulating levels of MCP-1 and TGF-β1 were increased in CAKUT adolescents compared with CAKUT children. In CAKUT children, levels of MCP-1 and TGF-β1 increased as CKD progressed, and MCP-1 and TGF-β1 were negatively and significantly correlated with eGFR and positively with urine protein. MCP-1 and TGF-β1 may contribute to the early detection of CKD and disease stage/progression in CAKUT children.

Platelets modulate cardiac remodeling via the collagen receptor GPVI after acute myocardial infarction.

Platelets play an important role in cardiovascular diseases. After acute myocardial infarction, platelets display enhanced activation and migrate into the infarct zone. Furthermore, platelets trigger acute inflammation and cardiac remodeling leading to alterations in scar formation and cardiac function as observed in thrombocytopenic mice. GPVI is the major collagen receptor in platelets and important for platelet activation and thrombus formation and stability. Antibody induced deletion of GPVI at the platelet surface or treatment of mice with recombinant GPVI-Fc results in reduced inflammation and decreased infarct size in a mouse model of AMI. However, the role of GPVI has not been fully clarified to date. In this study, we found that GPVI is not involved in the inflammatory response in experimental AMI using GPVI deficient mice that were analyzed in a closed-chest model. However, reduced platelet activation in response to GPVI and PAR4 receptor stimulation resulted in reduced pro-coagulant activity leading to improved cardiac remodeling. In detail, GPVI deficiency in mice led to reduced TGF-β plasma levels and decreased expression of genes involved in cardiac remodeling such as Col1a1, Col3a1, periostin and Cthrc1 7 days post AMI. Consequently, collagen quality of the scar shifted to more tight and less fine collagen leading to improved scar formation and cardiac function in GPVI deficient mice at 21d post AMI. Taken together, this study identifies GPVI as a major regulator of platelet-induced cardiac remodeling and supports the potential relevance of GPVI as therapeutic target to reduce ischemia reperfusion injury and to improve cardiac healing.

TGF-β1 overexpression in severe COVID-19 survivors and its implications for early-phase fibrotic abnormalities and long-term functional impairment.

In post-COVID survivors, transforming growth factor-beta-1 (TGF-β1) might mediate fibroblast activation, resulting in persistent fibrosis. In this study, 82 survivors of COVID-19-associated ARDS were examined at 6- and 24-months post-ICU discharge. At 6-months, quantitative CT analysis of lung attenuation was performed and active TGF-β1 was measured in blood and exhaled breath condensate (EBC). At 6-months of ICU-discharge, patients with reduced DmCO/alveolar volume ratio exhibited higher plasma and EBC levels of active TGF-β1. Plasma TGF-β1 levels were elevated in dyspneic survivors and directly related to the high-attenuation lung volume. In vitro, plasma and EBC from survivors induced profibrotic changes in human primary fibroblasts in a TGF-β receptor-dependent manner. Finally, at 6-months, plasma and EBC active TGF-β1 levels discriminated patients who, 24-months post-ICU-discharge, developed gas exchange impairment. TGF-β1 pathway plays a pivotal role in the early-phase fibrotic abnormalities in COVID-19-induced ARDS survivors, with significant implications for long-term functional impairment.

Significance of plasma TGF-b1 level detection in patients with T2DM with heart failure

Significance of plasma TGF-b1 level detection in patients with T2DM with heart failure

PRODUCTION OF GROWTH FACTORS AND DESQUAMATION OF ENDOTHELIOCYTES IN THE HEART IN ISCHEMIC CARDIOMYOPATHY

Highlights Dysregulation of angiogenesis may be the pathogenetic factor of ischemic cardiomyopathy (ICMP). Aim. To determine the content of growth factors and desquamated endothelial cells (DEC) in the blood from the coronary sinus and ulnar vein in association with the number of progenitor endothelial cells (PEC) in the blood from the ulnar vein in patients with coronary heart disease (CHD), suffering and not suffering from ICMP.Methods. The study included 30 patients with ICMР and 22 patients with CHD, and 18 healthy donors. The content of DEC (CD45–CD146+) was determined in blood from the cubital vein (peripheral) and coronary sinus, and the content of DEC (CD14+CD34+VEGFR2+) was determined in peripheral blood by flow cytometry (antibodies “BD Biosciences”, USA). The concentrations of VEGF-A, VEGF-B, PDGF, SDF-1, SCF, FGF-1, TGF-β1 in blood plasma from both locations were evaluated by multiplex analysis (set “Cloud-Clone Corp.”, USA).Results. The content of DEC in peripheral blood was elevated in patients with CHD of both groups, and in patients with ICMP in sinus blood was higher than in peripheral. At the same time, in patients with CHD without cardiomyopathy, an excess of PEC and SDF-1 in the blood from the ulnar vein was established in combination with an increase in the concentration of PDGF and a decrease in the content of VEGF-B in the blood from the coronary sinus relative to the parameters of systemic blood flow. In patients with ICMP, these changes were not detected, but there was an increase in the concentration of TGF-β1 in sinus blood compared with peripheral blood. Regardless of the presence of ICMP, the concentration of SCF, FGF-1, VEGF-A in the blood from the ulnar vein corresponded to the norm and that in sinus blood; the content of VEGF-A in the coronary bloodstream exceeded its systemic level.Conclusion. In patients with ICMP, desquamation of the coronary vascular endothelium is enhanced against the background of violations of its repair processes due to insufficient (relative to CHD without cardiomyopathy) mobilization of PEC from the bone marrow due to the absence of an excess of SDF-1 in the blood and their insufficient homing into the myocardium due to weak PDGF production in the heart.

Plasma Transforming Growth Factor-β1 (TGF-β1) Levels Increase with Age and Aortic Stenosis Progression and Low-Dose Galunisertib Attenuates Disease Progression in Mice

Objective: Aortic stenosis (AS) is an age-related degenerative valvular disease that ultimately leads to heart failure and sudden death. Platelet-derived TGF-β1 is a major source of plasma TGF-β1 and contributes to AS progression in LDLR -/-ApoB 100/100(LA100), a mouse model that spontaneously develops AS that simulates human pathology, including fibrosis and high shear gradient. Although risk factors, such as hypercholesteremia and hyperglycemia, are like those for coronary artery disease, lowering cholesterol and glucose has not been successful in stopping progression, and there is no biomarker for early detection of AS. In this study, we measured plasma TGF-β1 levels as a function of age in LA100 mice and human AS patients and tested a new approach of partial inhibition of TGF-β signaling to inhibit AS progression in LA100 mice, while limiting cardiotoxicity. Methods: We measured plasma TGF-β1 levels in controls, severe AS patients, and LA100 mice before they developed AS (2-months of age), when they developed moderate AS (1-year), and when they developed severe AS (2-years). LA100 mice at different stages of AS were treated with low-dose galunisertib, an inhibitor of the TGF-β receptor, ALK5. Results: Plasma levels of total TGF-β1 were significantly higher in AS patients (n=34) compared to healthy controls (n=22) and patient controls (n=47) (p&amp;lt;0.001 for AS patients vs. healthy controls and p=0.049 for AS patients vs. patient controls). Age was positively correlated with total TGF-β1 levels in AS patients (r=0.475; p=0.005), but not patient or healthy controls (r=0.12, p=0.43). LA100mice developed AS with aging and plasma TGF-β1 levels in LA100mice (n=21-52; ages 2 months-2 years) were significantly higher than those in age-matched WT mice (n=23); (p&amp;lt;0.0001). Plasma TGF-β1 levels correlated with age in LA100 mice (R=0.6, p=&amp;lt;0.0001). Low-dose oral galunisertib was administered at a concentration of 1 µM in drinking water, giving an average daily dose of ~0.06 mg/kg, compared to a human cancer treatment dose of ~4.2 mg/kg daily. Galunisertib treatment: a) halted AS progression at all stages in LA100 mice compared to vehicle-treated control mice (p&amp;lt;0.05, n=9-13), b) did not change plasma TGF-β1 levels, c) inhibited TGF-β1 signaling as shown by decreased Smad2 phosphorylation in valve cells compared to vehicle-treated animals. An age-dependent increase in neutrophil to lymphocyte ratio (NY/LY), a marker of inflammation, was observed in LA100 mice compared to control mice (n=12-31; p&amp;lt;0.01), and the NY/LY ratio correlated with total TGF-β1 plasma levels (R=0.16; p=0.020). Galunisertib treatment reduced NY/LY ratios in LA100 mice (p=0.03). Conclusions: 1. Increases in plasma TGF-β1 levels may be a valuable surrogate marker of AS progression. 2. Low-dose galunisertib attenuates AS progression in the LA100 mouse model without producing the cardiotoxicity associated with higher doses and thus may warrant further study in human AS patients.

Efficacy, safety, and correlative biomarkers of bintrafusp alfa in recurrent or metastatic nasopharyngeal cancer patients: a phase II clinical trial.

The strategy of dual blockade of TGF-β and PD-L1 pathways has not been previously tested in platinum-refractory recurrent or metastatic nasopharyngeal cancer (R/M NPC) patients. This study aimed to evaluate the safety and efficacy of bintrafusp alfa in refractory R/M NPC patients. In this single-arm, single-centre phase II clinical trial, 38 histologically confirmed R/M NPC patients were enrolled and administered with bintrafusp alfa every 2 weeks. Primary endpoint was objective response rate (ORR) per Response Evaluation Criteria in Solid Tumors version 1.1 (RECIST v1.1). Secondary endpoints included progression-free survival (PFS), overall survival (OS), duration of response (DOR), and safety. Thirty-eight patients were accrued (33 men; median age, 54 years). ORR was 23.7% (complete response, n=2; partial response, n=7). The median DOR was 19.2 months, median PFS was 2.3 months, median OS was 17.0 months, and 1-year OS rate was 63.2%. Unfortunately, 25 patients (65.7%) progressed within 8 weeks of treatment, 15 patients (39.5%) and 8 patients (21.1%) developed hyper-progressive disease (HPD) per RECIST v1.1 and tumor growth rate (TGR) ratio respectively. Sixteen patients (42.4%) experienced≥grade 3 treatment-related adverse events (TRAEs), most commonly anemia (n=9, 23.7%) and secondary malignancies (n=4, 10.5%). TRAEs led to permanent treatment discontinuation in 7 patients. Patients with strong suppression of plasma TGFβ1 level at week 8 were unexpectedly associated with worse ORR (9.1% vs 44.4%, P=0.046) and development of HPD. There was no correlation between PD-L1 expression and ORR. Bintrafusp alfa demonstrated modest activity in R/M NPC but high rates of HPD and treatment discontinuation secondary to TRAEs are concerning. The project was supported by Alice Ho Miu Ling Nethersole Charity Foundation Professorship Endowed Fund and Merck KGaA.

Amelioration of Tumor-promoting Microenvironment via Vascular Remodeling and CAF Suppression Using E7130: Biomarker Analysis by Multimodal Imaging Modalities.

E7130 is a novel anticancer agent created from total synthetic study of the natural compound norhalichondrin B. In addition to inhibiting microtubule dynamics, E7130 also ameliorates tumor-promoting aspects of the tumor microenvironment (TME) by suppressing cancer-associated fibroblasts (CAF) and promoting remodeling of tumor vasculature. Here, we demonstrate TME amelioration by E7130 using multi-imaging modalities, including multiplexed mass cytometry [cytometry by time-of-flight (CyTOF)] analysis, multiplex IHC analysis, and MRI. Experimental solid tumors characterized by large numbers of CAFs in TME were treated with E7130. E7130 suppressed LAP-TGFβ1 production, a precursor of TGFβ1, in CAFs but not in cancer cells; an effect that was accompanied by a reduction of circulating TGFβ1 in plasma. To our best knowledge, this is the first report to show a reduction of TGFβ1 production in TME. Furthermore, multiplex IHC analysis revealed reduced cellularity and increased TUNEL-positive apoptotic cells in E7130-treated xenografts. Increased microvessel density (MVD) and collagen IV (Col IV), an extracellular matrix (ECM) component associated with endothelial cells, were also observed in the TME, and plasma Col IV levels were also increased by E7130 treatment. MRI revealed increased accumulation of a contrast agent in xenografts. Moreover, diffusion-weighted MRI after E7130 treatment indicated reduction of tumor cellularity and interstitial fluid pressure. Overall, our findings strongly support the mechanism of action that E7130 alters the TME in therapeutically beneficial ways. Importantly, from a translational perspective, our data demonstrated MRI as a noninvasive biomarker to detect TME amelioration by E7130, supported by consistent changes in plasma biomarkers.

Resolution of fibrosis in mdx dystrophic mouse after oral consumption of N-163 strain of Aureobasidium pullulans produced β-glucan

Recent advances in the management of Duchenne muscular dystrophy (DMD), such as exon skipping and gene therapy, though have reached a clinical stage, the outcome at its best is still considered suboptimal. In this study, we evaluated a novel N-163 strain of Aureobasidium pullulans produced β-glucan (Neu-REFIX) for its potential as an adjuvant to slow down the progression of the disease by anti-inflammatory and anti-fibrotic effects. In this study, 45 mice in the three groups, 15 each in a group; Gr. 1 normal mice, Gr.2 mdx mice as vehicle, and Gr.3 mdx mice administered the N-163 β-glucan for 45 days. The N-163 β-glucan group showed a significant decrease in the plasma ALT, AST, and LDH levels (126 ± 69 U/l, 634 ± 371 U/l, 3335 ± 1258 U/l) compared with the vehicle group (177 ± 27 U/l, 912 ± 126 U/l, 4186 ± 398 U/l). Plasma TGF-β levels increased, and plasma IL-13 levels decreased in the N-163 group. The inflammation score of HE-stained muscle sections in the N-163 group (1.5 ± 0.8) was lower than that in the vehicle group (2.0 ± 0.8). The N-163 strain β-glucan group (24.22 ± 4.80) showed a significant decrease in the fibrosis area (Masson’s Trichrome-positive area) compared with the vehicle group (36.78 ± 5.74). The percentage of centrally nucleated fibres evaluated by Masson’s trichrome staining was 0 in the normal group, while it increased to 80% in the vehicle group but remained at 76.8% in the N-163 group. The N-163 β-glucan group showed a significant decrease in the fibrosis area. Considering their safety and easy oral consumption, Neu-REFIX β-glucan could be worth large multicentre clinical studies as adjuvant in slowing down the progress of DMD.

Abstract PO-095: TGF-β carrying exosomes in plasma of HNSCC as potential biomarkers of disease progression in patients with HNSCC

Abstract Background: The contribution of TGFβ+ exosomes, now referred to as small extracellular vesicles (sEV), in plasma of cancer patients to disease progression has been unclear. Levels of TGFβ in plasma often do not correlate with clinicopathological data, and TGFβ can exert dual (yin-yang) effects as a tumor promoter and a tumor inhibitor. We evaluated the potential of tumor-derived sEV dubbed TEX to serve as liquid tumor biopsy in patients with HNSCC. Methods: Plasma was obtained from 3 different cohorts of patients with HNSCC (n=97) and HDs (n=22). Clinicopathology/demographic data were available for all patients. sEV were isolated by ultrafiltration/size exclusion chromatography (SEC) from plasma or cell supernatants. TGF-β levels in plasma were assessed by ELISA; TGF-β expression in tissues was studied by IHC and immunoblots; immunoblots, flow cytometry and LC-MS/MS were used to measure TGF-β in isolated sEV. TGFB1 gene expression was analyzed using the TCGA Head and Neck Cancer database (n=520 cases and 44 NC). TGF-β signaling and activity of EVs were quantified in the reporter cell (MFB-F11) proliferation assays. Changes of TGF-β expression levels in plasma, tissues and sEV during carcinogenesis were studied in an orthotopic 4NQO mouse model. Results: Using the TCGA we found high expression levels of the vesiculation genes regulating EV production/release and of the TGFB1 gene expression in HNSCC. In the 4NQO orthotopic model of OSCC, cancer progression correlated with TGF-β accumulations in tumor tissues and a significant increase in circulating TGF-β+ sEV. In HNSCC patients, TGF-β levels and activity in plasma and TGF-β expression levels in tumor tissues were significantly elevated relative to controls but did not correlate with clinicopathologic data, tumor burden or disease outcome. TEX produced by HNSCC cell lines carried TGF-β and initiated Smad3 signaling in reporter cells. TGF-β inhibitors blocked this signaling. High TGFβ abundance in sEV was confirmed by proteomics and by RNAseq using the TCGA data base. Levels of TGF-β expression in sEV, sEV numbers and TGF-β activity of sEV were significantly elevated in HNSCC patients vs HDs’ sEV and correlated with tumor stage and LN involvement. Conclusions: TEX and sEV from HNSCC patients’ plasma carry TGF-β which reflects disease progression in HNSCC patients and in a mouse model of oral carcinogenesis. Neither plasma levels of TGF-β (ELISA) nor levels of TGF-β expression in the tumor (IHC) correlated with disease progression. Only sEV-associated TGF-β correlates with tumor burden and shows promise as a non-invasive biomarker of HNSCC progression and response to therapy. Citation Format: Nils Ludwig, Saigopalakrishna S. Yerneni, Theresa L. Whiteside. TGF-β carrying exosomes in plasma of HNSCC as potential biomarkers of disease progression in patients with HNSCC [abstract]. In: Proceedings of the AACR-AHNS Head and Neck Cancer Conference: Innovating through Basic, Clinical, and Translational Research; 2023 Jul 7-8; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2023;29(18_Suppl):Abstract nr PO-095.

The predictive value of plasma TGF-β levels in in-stent restenosis

Amaç: Stent-içi restenoz (SİR), stent implantasyonu sonrası erken ve geç dönem morbiditenin en önemli nedenlerinden birisidir. Biz bu çalışmada serum dönüştürücü büyüme faktörü-β (TGF-β) düzeyinin gelişebilecek SİR’i ön gördürüp gördürmeyeceğini araştırmayı planladık. Gereç ve Yöntem: Daha önce peruktan koroner girişim (PKG) uygulanmış ve herhangi bir endikasyonla koroner anjiografi (KAG) planlanmış toplam 82 hasta çalışmaya dahil edildi. SİR; stent içinde ≥%50 daralma olması şeklinde tanımlandı. 34 hastada restenoz saptanırken 48 hastada anjiografik olarak kritik lezyon saptanmadı. Tüm hastalarda plazma TGF- β düzeyi ölçüldü. Bulgular: SİR ile TGF- β düzeyi arasında istatistiksel olarak anlamlı bir ilişki saptanmadı (p=0,754). Yapılan alt grup çok değişkenli regresyon analizinde hedef damar çapı ile SİR arasında negatif, sağ koroner arter lezyonları ile SİR arasında pozitif korelasyon saptandı. Sonuç: Bu çalışma herhangi bir zamanda bakılan TGF-β’nın SİR öngördürücü bir özelliği olmadığını ortaya koymuştur