Lower VEGF Research Articles

A multicenter, prospective phase II trial of second-line aflibercept plus FOLFIRI in patients with metastatic colorectal cancer refractory to anti-EGFR agents (HGCSG1801): Updated analyses.

147 Background: The HGCSG1801 trial evaluated the treatment outcomes of aflibercept (AFL) plus FOLFIRI for patients(pts) with metastatic colorectal cancer (mCRC) refractory to an oxaliplatin-based regimen combined with an anti-EGFR agent. Here we report results of the updated efficacy, safety, and a biomarker analysis. Methods: This was a prospective open-label phase II trial. AFL (4 mg/kg iv) followed by FOLFIRI (irinotecan 180 mg/m 2 , leucovorin 200 mg/m 2 iv, bolus 5-fluorouracil [5-FU] 400 mg/m 2 , and infusional 5-FU 2400 mg/m 2 /46 h) was given every 2 weeks until progression or unacceptable toxicities. The primary endpoint was 6-month progression-free survival (PFS) rate, and the secondary endpoints included overall survival (OS), PFS, overall response rate (ORR), disease control rate (DCR), and adverse events. Angiogenic factors were analyzed in plasma sample using a Luminex multiplex assay using the Cox proportional hazards model. This study was sponsored by the Non-Profit Organization Hokkaido Gastrointestinal Cancer Study Group and supported by a grant from Sanofi. Results: Forty-three pts were enrolled between November 2019 and October 2022. The data cut-off date was April 30, 2024. The 6-month PFS rate was 59.0% (90% confidence interval [CI], 45.8–72.1%). Median PFS and OS were 7.3 months (95% CI, 5.5–11.0 months) and 18.8 months (95% CI, 12.9–26.6 months), respectively. The ORR was 20.9% (95% CI, 10.0–36.0%) and DCR was 88.4% (95% CI, 74.9–96.1%). No deaths and no new safety signals with a causal relation to the study treatment were observed. A biomarker analysis using pretreatment plasma samples showed a trend toward better PFS in pts with low VEGF-A (hazard ratio [HR] 0.40 [95% CI, 0.18-0.91]), TSP-2 (HR 0.40 [95% CI, 0.18-0.88]) or IL-8 levels (HR 0.43 [95% CI, 0.20-0.93]). There was also a trend toward better OS in pts with low levels of TSP-2 (HR 0.30 [95% CI, 0.11-0.81]) or TIMP-1 (HR 0.20 [95% CI, 0.06-0.69]). Conclusions: These updated data further support the activity and manageable safety profile of AFL plus FOLFIRI for pts with mCRC who failed an oxaliplatin-based regimen combined with an anti-EGFR agent. It was suggested the association between some angiogenic factors in plasma samples and the activity of AFL plus FOLFIRI in mCRC. Clinical trial information: jRCTs011190006 .

Histopathological growth patterns of gastric cancer liver metastasis and their association with patient prognosis.

484 Background: The histopathological growth patterns (HGPs) of liver metastasis reflect the complicated and varied interactions between tumor cells and the host microenvironment. We studied HGPs of gastric cancer liver metastasis (GCLM) and sought to predict them using radiomic and clinical variables. Methods: Hepatectomy was performed in 62 patients with GCLM at our university hospital between 2011 and 2021. HGPs were evaluated according to international consensus guidelines, and were classified as either desmoplastic HGP (dHGP) or non-dHGP. CD4, CD8, VEGF and CD31 were analyzed by immunohistochemistry. Eight hundred and fifty-one radiomics features were extracted from CT portal venous phase images and the optimal radiomics signature was determined by univariate analysis and LASSO regression. Multivariable regression analyses and ROC curves were used to assess the predictive performance of HGPs with a radiomic and clinical combined model. Results: Among 62 patients, 14 were categorized as dHGP, and 48 as non-dHGP. Non-dHGP vs. d-HGP was associated with higher tumor N stage and CEA level, and with decreased CD8 + T cells, and lower VEGF and CD31 expression. HGP status was independently associated with patient cancer-specific survival (CSS) (P＜0.05) multivariately.Analysis of percent relative contribution revealed that HGP ranked first for CSS (35.9%), ahead of T (28.9%) and N (28.9%) stage. A model with combined radiomic and clinical features demonstrated robust performance with area under the curve (AUC) values of 0.993 and 0.933 in the training and validation sets, respectively. Conclusions: Non-invasive determination of radiomic and clinical features was shown to predict HGPs types. Compared to dHGP, GCLM with non-dHGP exhibited significantly lower immune cell infiltration and decreased angiogenesis. Among features examined in GCLM, HGP was the most robust for prognostication. Univariate and multivariate analysis of prognostic factors for cancer-specific survival. Characteristics Univariate analysis Multivariate rergression analysis HR (95%CI) P HR (95%CI) P Gender ( Women vs Men ) 1.13(0.61-2.09) 0.71 Age ( >60years vs ≤60years) 1.08(0.60-1.70) 0.98 HGP ( non-desmoplastic HGP vs desmoplastic HGP ) 3.63(1.78-7.41) 0.00 3.48(1.37-8.88) 0.01 T stage ( T3-T4 vs T1-T2 ) 2.03(1.08-3.80) 0.03 2.29(1.15-4.58) 0.02 N stage ( N1-N3 vs N0 ) 3.46(1.66-7.23) 0.00 2.54(1.17-5.56) 0.02 CEA ( >5ng/ml vs ≤5ng/ml ) 2.37(1.25-4.48) 0.01 1.56(0.70-3.48) 0.27

VEGF-A as a prognostic indicator in advanced gastric and colorectal cancers and correlation with immune suppression involving myeloid-derived suppressor cells (MDSC).

250 Background: VEGF-A is important molecule should be controlled in cancer treatment since hypoxic and immunosuppressive conditions of tumor microenvironment are induced by VEGF-A. Myeloid-derived suppressor cells (MDSC), activated by chronic inflammation and VEGF-A, play critical roles in immunosuppression in patients with cancer. Methods: Serum levels of VEGF-A were measured by ELISA and circulating levels of MDSC were measured by Flow Cytometry (CD11b+, CD14-, CD33+). Patients with gastric and colorectal cancers were divided into two groups, high VEGF group (serum VEGF-A>330pg/ml) and lo group (≤330pg/ml). Results: Prognosis of stages III and IV gastric cancer with high VEGF-A was significantly worse than in low VEGF-A group, while difference was not significant in stages I and II. In colorectal cancer, the results were exactly same as found in gastric cancer. The levels of MDSC were significantly correlated with the levels of VEGF-A in patients with gastric and colorectal cancers. The levels of VEGF-A were also correlated with inflammatory markers including CRP and NLR (neutrophil to lymphocyte ratio), and inversely did to stimulation index of lymphocyte proliferation (SI: parameter of cellular immune reaction) and nutritional parameters such as albumin or rapid turnover protein. Conclusions: The observation in the present study suggest that VEGF-A is closely associated with inflammation-associated disease progression especially in advanced stages, and serve as useful marker of immunosuppression involving MDSC and prognosis in patients with gastric and colorectal cancers.

Vascular Endothelial Growth Factor Expression of Adipose-Derived Stromal Cells and Adipocytes Initiated from Fat Aspirations.

Fat grafting is frequently employed in aesthetic and reconstructive plastic surgery with a low complication rate. However, fat necrosis may occur in dependence of the mode of fat aspiration, processing of the tissue and graft size. Graft survival is critically dependent on the contained adipose-derived stromal cells (ADSCs), adipocyte precursors and their potential for vascular supply. This work investigated the potential role of the expression of vascular endothelial growth factor A (VEGF) and various cytokines by ADSCs and differentiated adipocytes as key factors of fat grafting. Adipokine expression of ADSCs and differentiated adipocytes were assessed using Proteome Profiler Arrays that detect 58 relevant proteins. Collected fat grafts could be categorized according to their adipokine expression into VEGFhigh and VEGFlow ADSCs groups, the former exhibiting higher content of VEGF-related angiopoietin-like 2, nidogen-1/entactin, CCL2/MCP-1 and elevated expression of IGFBPs in association with a fourfold higher VEGF expression. Differentiation of ADSCs into adipocytes increased VEGF concentrations in VEGFlow ADSCs but not in ADSCs exhibiting initial high VEGF concentrations. The adipocytes revealed high expression of HGF, leptin, CCL2/MCP-1, nidogen-1/entactin, M-CSF but lower induction of angiopoietin-like 2. Half of the ADSCs from fat grafts express high concentrations of VEGF and other adipokines that support angiogenesis and survival of this tissues following transfer. Differentiation of ADSClow cells to adipocytes may make up for the initially low VEGF expression, but this activation is 7-10 days delayed compared to the VEGFhigh ADSC cells and may fail to support angiogenesis from the beginning. This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Sex-specific associations between feto-placental growth and maternal physical activity volume and sitting time: Findings from the Queensland Family Cohort study.

Antenatal physical activity (PA) is associated with beneficial changes in placental growth and function; however, the effect of excessive sitting time is less clear. The aim of this study was to investigate whether feto-placental growth changes with maternal activity, and whether these associations differ in a sex-specific manner. This study included women enrolled in the Queensland Family Cohort study who self-reported PA and sitting time at 24 or 36 weeks of gestation. Placental growth factors and feto-placental growth parameters at delivery were analysed by PA volume and sitting time, as well as by fetal sex. Women who reported excessive sitting time during mid-pregnancy and had a female fetus showed higher placental PlGF (p=0.031) and FLT1 (p=0.032) mRNA expression with no difference in placental size at delivery. For the male, excessive sitting time during mid-pregnancy was associated with a lower placental weight (p=0.001) and placental surface area (p=0.012) and a higher birthweight to placental weight (BWPW) ratio (p=0.042), with no change in placental growth factors. Moderate volume PA during mid-pregnancy was associated with lower VEGFA mRNA expression in the male placenta (p=0.005) and a higher abdominal circumference in the female neonate (p=0.042), with no overall difference in placental weight or birthweight for either sex. The results of this study suggest that mid-pregnancy may be an important timepoint for programming of feto-placental growth in relation to maternal activity. Our findings highlight the independent benefits of reducing sitting time during pregnancy, particularly for women carrying male fetuses.

Visfatin and VEGF levels are not increased in adolescent girls with polycystic ovary syndrome.

PCOS is one of the most commonly occurring endocrinopathies among women and increasingly affects adolescent populations. The connection between PCOS and various endocrinological, psychological, and CVD is increasingly recognized. Some studies have shown elevated levels of visfatin and VEGF among patients with PCOS, which are markers of vascular endothelial dysfunction. In our study, we evaluated the concentration of these parameters, focusing solely on a group of adolescents with PCOS, to assess whether these early markers of CVD are present at an early stage of diagnosis. In total, 80 adolescent girls participated in the study. 47 adolescents diagnosed with PCOS were included in the study group (mean age 15.68 ± 1.18 years, BMI 26.66 ± 6.41 kg/m2), while the remaining 33 regularly menstruating individuals (mean age 15.79 ± 1.22 years, BMI 25.44 ± 7.24 kg/m2) were assigned to the control group. Each participant underwent imaging, biochemical, and hormonal tests. Additionally, markers of endothelial dysfunction: VEGF and visfatin, were measured in all adolescents. Both VEGF and visfatin levels did not differ significantly between PCOS and control group (p=0.30 and p=0.15, respectively). In the group of adolescent girls with PCOS, visfatin was significantly correlated with HDL, FSH, cortisol, and testosterone levels >55 ng/dl. VEGF was significantly correlated with fasting glucose, glucose levels after OGTT, estradiol, and waist circumference >80 cm. It can be indirectly inferred that both visfatin and VEGF should not be used as early markers for cardiometabolic complications among adolescent patients with PCOS. On the other hand, low visfatin levels, through their negative correlation with HDL, may have a protective effect on cardiovascular complications, while low VEGF levels, through their positive correlation with glucose levels, may have a protective influence on carbohydrate metabolism disorders.

Exploring the therapeutic applications of nano-therapy of encapsulated cisplatin and anthocyanin-loaded multiwalled carbon nanotubes coated with chitosan-conjugated folic acid in targeting breast and liver cancers

This study aimed to assess the targeted nano-therapy of encapsulated cisplatin (Cis) and anthocyanin (Ant)-loaded multiwalled carbon nanotubes (CNT) coated with chitosan conjugated folic acid on breast MCF7 and liver HepG2 cancer cells. Zeta potential, UV-spectroscopy, FTIR, TEM, and SEM were used to evaluate CNT, its modified form (CNT Mod), CNT-loaded Cis NPs, CNT-loaded Ant NPs, and CNT- Cis + Ant NPs. All treatments induced apoptosis-dependent cytotoxicity in both cell lines as revealed functionally by the MTT assay, morphologically (DNA degradation) by acridine orange/ethidium bromide (AO/EB) double staining, and molecularly (Bax upregulation and Bcl2 downregulation) by real-time PCR, with best effect for the combined treatment (CNT- Cis + Ant NPs). This combined treatment also significantly reduced inflammation (low TNFα), migration (low MMP9 and high TIMP1), and angiogenesis (low VEGF), while significantly increasing antioxidant status (high Nrf2 and OH-1) in MCF7 and HepG2 cells compared to other treatments. Interestingly, cells treated with CNT Mod exhibited higher cytotoxic, apoptotic, anti-migratory, and anti-angiogenic potentials relative to CNT-treated cells. In conclusion, targeted nano-therapy of encapsulated cisplatin and anthocyanin-loaded carbon nanotubes coated with chitosan conjugated folic acid can efficiently combat breast and liver cancers by sustained release, in addition to its apoptotic, antioxidant, anti-inflammatory, anti-metastatic, and anti-angiogenic effects.

ETHE1 dampens colorectal cancer angiogenesis by promoting TC45 Dephosphorylation of STAT3 to inhibit VEGF-A expression

Angiogenesis is critical for colorectal cancer (CRC) progression, but its mechanisms remain unclear. Here, we reveal that ethylmalonic encephalopathy protein 1 (ETHE1), an essential enzyme in hydrogen sulfide catabolism, inhibits VEGF-A expression and tumor angiogenesis in vitro and in vivo. Moreover, we find that this biological function of ETHE1 depends on the STAT3/VEGF-A pathway. Further investigation demonstrates that ETHE1 promotes the interaction between T cell protein tyrosine phosphatase (TC45) and STAT3, resulting in decreased STAT3 phosphorylation and inhibition of the STAT3 signaling pathway. In clinical samples, we find that ETHE1 is downregulated in CRC and positively correlates with survival outcomes of CRC patients. Meanwhile, the negative correlation of ETHE1 and VEGF-A expression is verified in CRC specimens, and the patients with low ETHE1 and high VEGF-A expression exhibits poorer prognosis. Collectively, our study identifies ETHE1 as a novel regulator of tumor angiogenesis, implying its potential as a prognostic biomarker and promising antiangiogenic target for CRC patients.

Placental growth factor mediates pathological uterine angiogenesis by activating the NFAT5-SGK1 signaling axis in the endometrium: implications for preeclampsia development

After menstruation the uterine spiral arteries are repaired through angiogenesis. This process is tightly regulated by the paracrine communication between endometrial stromal cells (EnSCs) and endothelial cells. Any molecular aberration in these processes can lead to complications in pregnancy including miscarriage or preeclampsia (PE). Placental growth factor (PlGF) is a known contributing factor for pathological angiogenesis but the mechanisms remain poorly understood. In this study, we investigated whether PlGF contributes to pathological uterine angiogenesis by disrupting EnSCs and endothelial paracrine communication. We observed that PlGF mediates a tonicity-independent activation of nuclear factor of activated T cells 5 (NFAT5) in EnSCs. NFAT5 activated downstream targets including SGK1, HIF-1α and VEGF-A. In depth characterization of PlGF - conditioned medium (CM) from EnSCs using mass spectrometry and ELISA methods revealed low VEGF-A and an abundance of extracellular matrix organization associated proteins. Secreted factors in PlGF-CM impeded normal angiogenic cues in endothelial cells (HUVECs) by downregulating Notch-VEGF signaling. Interestingly, PlGF-CM failed to support human placental (BeWo) cell invasion through HUVEC monolayer. Inhibition of SGK1 in EnSCs improved angiogenic effects in HUVECs and promoted BeWo invasion, revealing SGK1 as a key intermediate player modulating PlGF mediated anti-angiogenic signaling. Taken together, perturbed PlGF-NFAT5-SGK1 signaling in the endometrium can contribute to pathological uterine angiogenesis by negatively regulating EnSCs-endothelial crosstalk resulting in poor quality vessels in the uterine microenvironment. Taken together the signaling may impact on normal trophoblast invasion and thus placentation and, may be associated with an increased risk of complications such as PE.

Better clinical outcomes and lower triggering of inflammatory cytokines for allogeneic hematopoietic cell transplant recipients treated in home care versus hospital isolation - the Karolinska experience.

After allogeneic hematopoietic cell transplantation (Allo-HCT) and conditioning, patients are typically placed in isolated hospital rooms to prevent neutropenic infections. Since 1998, we've offered an alternative: home care for patients living within a one to two-hour drive of the hospital. In Sweden this approach includes daily visits by an experienced nurse and daily phone consultations with a unit physician. When necessary, patients receive transfusions, intravenous antibiotics, and total parenteral nutrition at home. Our initial study report compared 36 home care patients with 54 hospital-treated controls. Multivariate analysis found that home care patients were discharged earlier to outpatient clinics, required fewer days of total parenteral nutrition, had less acute graft-versus-host disease (GVHD) grade II-IV, and lower transplantation-related mortality (TRM) and lower costs. Long-term follow-up showed similar chronic GVHD and relapse rates in both groups, with improved survival rates in the home care group. A subsequent comparison of 146 home care patients with hospital-treated controls indicated that home care and longer home stays were associated with lower grades of acute GVHD. Home care was found to be safe and beneficial for children and adolescents. Over two decades, 252 patients received home care post-Allo-HCT without any fatalities at-home. Ten-year outcomes showed a 14% TRM and a 59% survival rate. In 2020, an independent center confirmed the reduced risk of acute GVHD grades II-IV for patients treated in home care. Here, we report for the first time that home care patients also demonstrate a less inflammatory systemic cytokine profile. We found higher levels of IFN-γ, IL-2, IL-5, IL-13, GM-CSF, and G-CSF, but lower VEGF in hospital-treated patients, which may contribute to acute GVHD grades II-IV. In conclusion, home-based treatment following Allo-HCT yields multiple promising clinical outcomes and improved systemic inflammatory markers, which may contribute to less development of life-threatening GVHD.

Donor plasma VEGF-A as a biomarker for myocardial injury and primary graft dysfunction after heart transplantation

BackgroundVascular endothelial growth factor (VEGF)-A is an angiogenic and proinflammatory cytokine with profound effects on microvascular permeability and vasodilation. Several processes may induce VEGF-A expression in brain-dead organ donors. However, it remains unclear whether donor VEGF-A is linked to adverse outcomes after heart transplantation. MethodsWe examined plasma VEGF-A levels from 83 heart transplant donors as well as the clinical data of these donors and their respective recipients operated between 2010 and 2016. The donor plasma was analyzed using Luminex-based Multiplex and confirmed with a single-target ELISA. Based on donor VEGF-A plasma levels, the recipients were divided into three equal-sized groups (low VEGF <500 ng/L, n=28; moderate VEGF 500-3000 ng/L, n=28; and high VEGF >3000 ng/L, n=27). Biochemical and clinical parameters of myocardial injury as well as heart transplant and kidney function were followed-up for one year, while rejection episodes, development of cardiac allograft vasculopathy, and mortality were monitored for five years. ResultsBaseline parameters were comparable between the donor groups, except for age, where median ages of 40, 45, and 50 were observed for low, moderate, and high donor plasma VEGF levels groups, respectively, and therefore donor age was included as a confounding factor. High donor plasma VEGF-A levels were associated with pronounced myocardial injury (TnT and TnI), a higher inotrope score, and a higher incidence of primary graft dysfunction in the recipient after heart transplantation. Furthermore, recipients with allografts from donors with high plasma VEGF-A levels had a longer length of stay in the intensive care unit and the hospital, and an increased likelihood for prolonged renal replacement therapy. ConclusionsOur findings suggest that elevated donor plasma VEGF-A levels were associated with adverse outcomes in heart transplant recipients, particularly in terms of myocardial injury, primary graft dysfunction, and long-term renal complications. Donor VEGF-A may serve as a potential biomarker for predicting these adverse outcomes and identifying extended donor criteria.

Differential Effects of n-3 and n-6 Polyunsaturated Fatty Acids on Placental and Embryonic Growth and Development in Diabetic Pregnant Mice.

The present study aimed to investigate the differential effects of n-3 and n-6 polyunsaturated fatty acids (PUFAs) on placental and embryonic development. Pregnant mice were assigned to five groups: healthy control (HC), diabetes mellitus control (DMC), diabetes + low-dose n-3 PUFA (Ln-3), diabetes + high-dose n-3 PUFA (Hn-3), and diabetes + n-6 PUFA (n-6). On E12.5d, the Hn-3 group, but not the n-6 group, had a higher placenta weight. The weight ratio of embryo to placenta in the n-6 group was significantly lower than in the Hn-3 group but higher than in the DMC group. The Hn-3 group had significantly higher protein levels of VEGF, IGF-1, and IGFBP3, while the n-6 group had lower VEGF than the DMC group. Compared with the DMC group, embryonic Cer-16:0 was significantly higher in the Hn-3 group, while embryonic PC (36:6), PC (38:7), and PE (40:7) were significantly lower in the n-6 group. The embryo and placenta weights were positively correlated with placental VEGF, IGFBP3, and embryonic Cer-16:0, and they were negatively correlated with embryonic PC (36:6) and PE (40:7). The weight ratio of embryo to placenta was negatively correlated with embryonic PC (36:6). In addition, embryonic Cer-16:0 was positively correlated with placental VEGF and IGFBP3. In conclusion, n-3 PUFA and n-6 PUFA improved placental and embryonic growth through different mechanisms.

Immunoexpression of CD44, p16 and VEGF in oral cancer.

The aim of the present study was to examine the immunoexpression of CD44, p16 and VEGF in oral squamous cell carcinoma (OSCC) and correlate them to clinicopathological parameters and survival outcomes in order to clarify their prognostic impact. A total of 68 individuals with OSCC recruited between 2011 and 2015 from two referral centres were enrolled in the study. The samples were placed on silanized glass slides and subjected to immunohistochemistry using anti-p16, anti-CD44 and anti-VEGF antibodies. The H Score was used for p16 and VEGF, while CD44 was scored according to the percentage of stained cells. Chi-square tests and Fisher's exact probability tests were used to compare clinicopathological characteristics according to the immunohistochemical expression, while overall survival and disease-free survival were estimated and compared using the Kaplan-Meier method and log-rank test, respectively. For all hypothesis tests, the level of significance was set at P ≤ 0.05. No correlation was observed between the expression of tumour VEGF, p16 and CD44, and the clinicopathological characteristics analysed. Patients with high stromal VEGF expression had better disease-free survival than patients with low VEGF expression (P = 0.023). In summary, P16, CD44 and tumour VEGF did not prove to be good prognostic biomarkers. Stromal VEGF expression is suggested to be a good candidate prognostic biomarker, although additional studies are needed.

The effect of leizumab on serum vascular endothelial growth factor, IL-6, MCP-1 inflammatory factors in neovascular glaucoma.

This study aimed to assess Leizumab's effect on serum endothelial growth factor, interleukin-6 (IL-6), monocyte chemotactic protein-1 (MCP-1), and inflammatory factors in neovascular glaucoma patients. 80 eligible patients treated between January 2021 and April 2023 were enrolled, randomly divided into control and study groups. The control group underwent vitrectomy while the study group received preoperative intravitreal rituximab injection. Measurements included serum and aqueous humor VEGF/PEDF, IL-6/MCP-1 levels, postoperative rebleeding/retinal detachment, and visual acuity changes over 6 weeks. After surgery, patients showed reduced serum VEGF/PEDF levels (P < 0.05), with decreased VEGF and increased PEDF in aqueous humor (P < 0.05). The study group had lower VEGF and higher PEDF levels than the control (P < 0.05). Serum IL-6/MCP-1 levels reduced post-surgery, with the study group lower than control (P < 0.05). Intraocular rebleeding was lower in the study group (P < 0.05), while retinal detachment rates were similar (P > 0.05). Visual acuity differed significantly from week 1 to 6 post-surgery (P < 0.05), with higher acuity in the study group during weeks 1-4 (P < 0.05). Weeks 5-6 follow-up showed no significant difference (P > 0.05). Pre-vitrectomy ranibizumab injection effectively reduced bleeding, VEGF/PEDF levels, inflammatory factors, and improved visual recovery.

Plasma VEGFA and PGF impact longitudinal tau and cognition in preclinical Alzheimer's disease.

Vascular dysfunction is increasingly recognized as an important contributor to the pathogenesis of Alzheimer's disease. Alterations in vascular endothelial growth factor (VEGF) pathways have been implicated as potential mechanisms. However, the specific impact of VEGF proteins in preclinical Alzheimer's disease and their relationships with other Alzheimer's disease and vascular pathologies during this critical early period remain to be elucidated. We included 317 older adults from the Harvard Aging Brain Study, a cohort of individuals who were cognitively unimpaired at baseline and followed longitudinally for up to 12 years. Baseline VEGF family protein levels (VEGFA, VEGFC, VEGFD, PGF and FLT1) were measured in fasting plasma using high-sensitivity immunoassays. Using linear mixed effects models, we examined the interactive effects of baseline plasma VEGF proteins and amyloid PET burden (Pittsburgh Compound-B) on longitudinal cognition (Preclinical Alzheimer Cognitive Composite-5). We further investigated if effects on cognition were mediated by early neocortical tau accumulation (flortaucipir PET burden in the inferior temporal cortex) or hippocampal atrophy. Lastly, we examined the impact of adjusting for baseline cardiovascular risk score or white matter hyperintensity volume. Baseline plasma VEGFA and PGF each showed a significant interaction with amyloid burden on prospective cognitive decline. Specifically, low VEGFA and high PGF were associated with greater cognitive decline in individuals with elevated amyloid, i.e. those on the Alzheimer's disease continuum. Concordantly, low VEGFA and high PGF were associated with accelerated longitudinal tau accumulation in those with elevated amyloid. Moderated mediation analyses confirmed that accelerated tau accumulation fully mediated the effects of low VEGFA and partially mediated (31%) the effects of high PGF on faster amyloid-related cognitive decline. The effects of VEGFA and PGF on tau and cognition remained significant after adjusting for cardiovascular risk score or white matter hyperintensity volume. There were concordant but non-significant associations with longitudinal hippocampal atrophy. Together, our findings implicate low VEGFA and high PGF in accelerating early neocortical tau pathology and cognitive decline in preclinical Alzheimer's disease. Additionally, our results underscore the potential of these minimally-invasive plasma biomarkers to inform the risk of Alzheimer's disease progression in the preclinical population. Importantly, VEGFA and PGF appear to capture distinct effects from vascular risks and cerebrovascular injury. This highlights their potential as new therapeutic targets, in combination with anti-amyloid and traditional vascular risk reduction therapies, to slow the trajectory of preclinical Alzheimer's disease and delay or prevent the onset of cognitive decline.

Method of comprehensive physiotherapy of dental patients after COVID-19

To describe the mechanism of action and clinical effectiveness of the developed combined physiotherapy method, including the induced technique of piracetam iontophoresis on the frontooccipital technic and acupuncture laser therapy in dental patients with complaints of edentulism progression after COVID-19 on the basis of the analysis of single studies on the post-COVID loss of teeth treatment. A number of patients equal 120 who complained of tooth loss after COVID-19 during the past 6 months were examined. The following initial and end points were considered: dental bleeding and inflammation scores, vascular and endothelial dysfunction markers - levels of intercellular adhesion molecules and their receptors (SlCAM-1, SVCAM-1, VEGF-A, ET-1) before and after treatment. Negative correlation between VEGF-A (pg/ml) concentration in peripheral blood serum and sVCAM-1 (ng/ml) level in the examined patients (r=0.4830, p<0.05) and strong inverse correlation between slCAM-1 (ng/ml) level and sVCAM-1 (r=0.7696, p<0.01) have been established. More significant effects after application of the combined induced method on the head's structures and laser acupuncture have been noted than after acupuncture laser exposure and after inducing technique separately, namely in the form of dental inflammation score correction by 1.76 times (p<0.001), decrease of bleeding score by 2.6 (p<0.05), decrease of concentration of SVCAM-1 by 1.7 times and SlCAM-1 by 2 times (p<0.001), increase of endothelin level by 1.7 times as well as the initial low VEGF-A (pg/ml) by 1.5 times (p<0.01). The developed physiotherapeutic complex, which includes laser acupuncture physiotherapy and induced technique of 5% piracetam iontophoresis, can potentially be considered as a physioprophylactic and therapeutic model of post-COVID edentulism.

BDNF/TrkB signaling in stable coronary artery disease

Aim. To study the serum content of brain-derived neurotrophic factor (BDNF) and tyrosine kinase receptor B (TrkB) in patients with coronary artery disease (CAD) and evaluate the relationship of BDNF/TrkB signaling with the severity of coronary atherosclerosis, systemic inflammation (IL-2, IL-4, IL-6, IL-10, TNF-α) and angiogenesis (VEGF).Material and methods. The study included 99 patients with stable CAD who underwent coronary angiography and 30 healthy volunteers. Coronary atherosclerosis was assessed using the Gensini score (GS). In blood serum, the concentrations of BDNF, TrkB, VEGF, IL-2, IL-4, IL-6, IL-10, TNF-α were determined using the enzyme immunoassay. Cluster, correlation, and regression analyzes were used.Results. In patients with CAD, a wide range of variations in BDNF concentrations was observed. To determine homogeneous groups using the k-means clustering, three clusters with different BDNF/TrkB axis vectors were identified. Patients differed in the severity of coronary atherosclerosis, the manifestation of the inflammatory reaction, and the intensity of angiogenesis. In patients with initial and moderate atherosclerotic changes in the coronary arteries, a normal concentration of BDNF and an increased level of TrkB (22,35/1,18 ng/ml) were noted. In patients with severe coronary atherosclerosis, two different BDNF/TrkB variants have been identified. Decreased BDNF and increased TrkB (6,0/1,52 ng/ml) were associated with low VEGF and increased IL-6. Elevated BDNF and normal TrkB values (26,95/0,96 ng/ml) were characteristic of patients with high VEGF expression, indicating angiogenesis activation and/or vulnerable plaques. A direct relationship between BDNF and VEGF (r=0,536, p&lt;0,001) and an inverse relationship with TrkB (r=-0,301, p=0,019), IL-6 (r=-0,306, p=0,002) was revealed. TrkB levels were correlated with TNF-α (r=0,403, p=0,001). Regression analysis showed that BDNF expression is influenced by TrkB (β=-0,237, p=0,009), VEGF (β=0,490, p&lt;0,001), IL-6 (β=-0,339, p&lt;0,001).Conclusion. In patients with stable CAD, different levels of BDNF/TrkB expression were found, which were associated with coronary atherosclerosis severity. BDNF/TrkB signaling is involved in the regulation of inflammation and angiogenesis in stable CAD.

Metformin Regulates the miR-205/VEGFA Axis in Renal Cell Carcinoma Cells: Exploring a Clinical Synergism with Tyrosine Kinase Inhibitors

Introduction: Metformin (MF) intake could be associated with a favorable outcome in sunitinib (SUT)- and axitinib (AX)-treated clear cell renal cell carcinoma (ccRCC) patients. Functionally, MF induces miR-205, a microRNA serving as a tumor suppressor in several cancers. Methods: Real-time quantitative PCR, viability assays, and Western blotting analyzed MF and SUT/AX effects in RCC4 and 786-O cells. A tetracycline-inducible overexpression model was used to study the role of miR-205 and its known target gene, VEGFA. We analyzed miR-205 and VEGFA within a public and an in-house ccRCC cohort. Human umbilical vein endothelial cell (HUVEC) sprouting assays examined miR-205 effects on angiogenesis initiation. To determine the influence of the von Hippel-Lindau tumor suppressor (VHL), we examined VHL^wt reexpressing RCC4 and 786-O cells. Results: Viability assays confirmed a sensitizing effect of MF toward SUT/AX in RCC4 and 786-O cells. Overexpression of miR-205 diminished VEGFA expression – as did treatment with MF. Tumor tissue displayed a downregulation of miR-205 and an upregulation of VEGFA. Accordingly, miR-205 caused less and shorter vessel sprouts in HUVEC assays. Finally, VHL^wt-expressing RCC4 and 786-O cells displayed higher miR-205 and lower VEGFA levels. Conclusion: Our results support the protective role of MF in ccRCC and offer functional insights into the clinical synergism with tyrosine kinase inhibitors.

Mass Spectrometry Based Detection of Naturally Expressed FVIII in Liver Sinusoidal-like Endothelial Cells from Healthy Donor and HA Patients with Nonsense Mutations

MS Background: Liver sinusoidal endothelial cells (LSEC) are unique endothelial cells and secretion and expression of FVIII is one of their key functions. Pluripotent stem cells from healthy donors can be a source of generating organ-specific endothelial cells in vitro. However, maintaining the specification of LSEC markers remains a challenge. Currently, EBs with a combination of AMP activation,TGF-ß inhibition,and hypoxia are a state of the art to derive LSEC-like EC from IPS. A 2D monolayer-based approach shows potential in culturing these ECs and in maintaining their correct specification (Fig. 1).IPSc-differentiated LSEC were analyzed for the presence of LSEC-specific markers FCGR2b, Stab2 &amp; F8 by rtPCR. Our data shows that day 10 angioblasts mark an increase of venous markers COUP-TF and NT5E under low VEGF-A conditions. Upon LSEC induction LSEC markers are upregulated compared to HUVEC:FCGR2b(2x), Stab2(37x), and F8(7x) (not shown). Flow cytometry(FC) analyses depict a KDR +CD56 + phenotype for mesoderm on day 4 ,a venous CD31 +CD34 +CD184 lowCD73 high phenotype for day 6 angioblast and a CD34 +FCGR2b +Stab2 +LYVE1 + LSEC-LC population successfully maintained until Day 15 (not shown).ELISA measurements (Fig. 2) using a FVIII-A3 specific biotinylated nanobody detects FVIII in LSEC-LC from healthy donor, HA-patient with I22I and in both high risk inhibitor nonsense mutations R1960X and R2228X. These preliminary results warranted the confirmation of detection as well as the need to decipher the differences in the FVIII protein among the healthy donor and patients with HA. Aim: Detection of endogenous FVIII protein in healthy donors and patients using immunofluorescence(IF). Detection of FVIII protein using mass spectrometry (MS) Methods: IF was conducted using a monoclonal antibody (GMA8021) and the difference in protein levels was measured using the mean fluorescence intensity (MFI) values. An immunoprecipitation-based method to detect the natural FVIII protein using MS was developed. Initially, an ultracentrifugation step eliminated the background proteome which allowed for the antibody (SAF8C) based capture of FVIII using a column-based elution technique. The eluted sample was trypsinized and detected via nano-LC coupled to LC/MS Orbitrap. Results: The MFI of the healthy donor is 2.8x, 2.5x, 2.31x, 2x and 1.3x higher than F8KO, HUVEC, HA-patient samples R2228X, R1960X and I22I,respectively. Healthy donor shows the presence of 5 FVIII peptides. Patient sample with mutation I22I shows the presence of 1 FVIII peptide. Patient sample with nonsense mutation R1960X showed 2 FVIII peptides while patient sample with R2228X was unable to show any FVIII peptides. Conclusion: While FVIII detection in multiple overexpressed models of HA have been established, our current LSEC model is a milestone in detecting the natural, native state of the FVIII protein. This LSEC-LC model provides a 7-fold higher F8 expression compared to HUVEC in vitro system. FC analysis shows the right phenotype for modeling IPSc into LSEC. The additional presence of LSEC-specific receptors and confirmation of FVIII in the healthy donor and patients with nonsense mutations enables a more native and precise disease modeling for HA research and therapy.

Utilizing the Systemic Immune-Inflammation Index and Blood-Based Biomarkers in Association with Treatment Responsiveness amongst Patients with Treatment-Resistant Bipolar Depression.

(1) Background: Inflammation is associated with depressive illness and treatment resistance. This study assessed a novel inflammatory index, the Systemic Immune-Inflammation Index (SII), in patients diagnosed with treatment-resistant bipolar depression (TRBDD) before and after treatment with escitalopram (ESC) and celecoxib (CBX) add-on or ESC and placebo (PBO), and compared them to healthy control (HC) subjects. (2) Methods: This is a secondary biological analysis from a double-blind randomized placebo-controlled trial of CBX augmentation in TRBDD. Our subsample with available complete blood count (CBC) data included 52 TRBDD subjects, randomized into an ESC + CBX, (n = 29), an ESC + PBO arm (n = 23), and an HC group (n = 32). SII was calculated from the CBC with differential (SII = platelets x neutrophils/lymphocytes) at baseline and end of treatment (8 weeks). Blood inflammation biomarkers, growth factors, and kynurenine metabolites were determined at both timepoints. Depressive symptom severity was the primary outcome, using the HAMD-17 rating scale score to quantitate treatment response and remission rates. (3) Results: Baseline SII did not discriminate TRBDD from HC, nor was it associated with HAMD-17 score at any timepoint, although it was significantly associated with lower baseline VEGF (p = 0.011) and higher week 8 levels of IL1-β (p = 0.03) and CRP (p = 0.048). Post-treatment HAMD-17 was not independently predicted using baseline SII unless an interaction with age was present (p = 0.003 was included), even after relevant adjustments. A similar effect was seen with baseline neutrophils. (4) Conclusions: While SII was not an independent predictor of treatment outcome, elevated baseline SII was a predictor of poor treatment response amongst older patients with TRBDD.