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Heterogenous expression of CD31 in human samples in situ and patient-derived angiosarcoma cells in vitro. A, CD31 IHC staining in an epitheloid human angiosarcoma showing very low expression in this tumor area ( Ã ), whereas neoplastic endothelial cells forming a vascular structure are strongly positive (arrowheads). Other regions of the tumor showed strong expression of CD31. See also Supplementary Fig. S1A for more CD31 stainings. B, Immunofluorescence double staining of the same case showing multiple ERG-positive tumor cell nuclei (green). Few cells show membranous coexpression of CD31 (CD31 high , red staining), whereas most cells in this part of the tumor were CD31 low. Arrowheads highlight regions with CD31/ERG colocalization, while ( Ã ) marks ERG positive regions with low CD31 expression. Immunoblot analysis (C) and densitometric quantification (D) of CD31 expression in normal primary endothelial cells (HUVEC and HDMEC), the hybrid cell line EA.hy926 (HUVEC fused with lung adenocarcinoma cells), and angiosarcoma (AS) cell lines ISO-HASc.1, HAMON and ASM, showing lower CD31 levels in neoplastic endothelial cells (n ¼ 3). E, FACS analysis using a CD31-PE or isotype-PE (control) antibody was performed in all indicated cell lines. Although non-neoplastic primary endothelial cells (HUVEC and HDMEC) presented homogenous CD31 expression, the EA.hy926 cell line as well as the angiosarcoma cell lines ISO-HASc.1 and ASM showed a substantial proportion of CD31 low cells. See also Supplementary Fig. S1B for a representative CD31 FACS analysis of the HAMON angiosarcoma cell line and Supplementary Fig. S1C for statistical analysis of all cell lines.
Source publication
Purpose:
Angiosarcomas (AS) are soft tissue sarcomas with endothelial differentiation and vasoformative capacity. Most AS show strong constitutive expression of the endothelial adhesion receptor CD31/PECAM-1 pointing to an important role of this molecule. However, the biological function of CD31 in AS is unknown.
Experimental design:
The express...
Citations
... Moreover, despite recent studies into combination therapies in the management of AS, response to first-line systemic therapy is poor because of the absence of predictive biomarkers and significant intra-tumor heterogeneity [26]. Thus far, apart from one study that has employed a morphological approach to identify a potential marker of chemoresistance in AS, that is CD31 low cells, which relies on enhanced YAP signaling to improve redox status and is doxorubicin-resistant in AS [27], to our knowledge, this is the first study to identify the potential biomarkers of chemoresistance in AS via an omics approach. ...
Angiosarcomas, clinically aggressive cancers of endothelial origin, are a rare subtype of soft-tissue sarcomas characterized by resistance to chemotherapy and dismal prognosis. In this study, we aim to identify the transcriptomic biomarkers of chemoresistance in angiosarcoma. We examined 72 cases of Asian angiosarcomas, including 35 cases treated with palliative chemotherapy, integrating information from NanoString gene expression profiling, whole transcriptome profiling (RNA-seq), immunohistochemistry, cell line assays, and clinicopathological data. In the chemoresistant cohort (defined as stable disease or progression), we observed the significant overexpression of genes, including SPP1 (log2foldchange 3.49, adj. p = 0.0112), CXCL13, CD48, and CLEC5A, accompanied by the significant enrichment of myeloid compartment and cytokine and chemokine signaling pathways, as well as neutrophils and macrophages. RNA-seq data revealed higher SPP1 expression (p = 0.0008) in tumor tissues over adjacent normal compartments. Immunohistochemistry showed a significant moderate positive correlation between SPP1 protein and gene expression (r = 0.7016; p < 0.00110), while higher SPP1 protein expression correlated with lower chemotherapeutic sensitivity in patient-derived angiosarcoma cell lines MOLAS and ISOHAS. In addition, SPP1 mRNA overexpression positively correlated with epithelioid histology (p = 0.007), higher tumor grade (p = 0.0023), non-head and neck location (p = 0.0576), and poorer overall survival outcomes (HR 1.84, 95% CI 1.07–3.18, p = 0.0288). There was no association with tumor mutational burden, tumor inflammation signature, the presence of human herpesvirus-7, ultraviolet exposure signature, and metastatic state at diagnosis. In conclusion, SPP1 overexpression may be a biomarker of chemoresistance and poor prognosis in angiosarcoma. Further investigation is needed to uncover the precise roles and underlying mechanisms of SPP1.
... The Western blotting results showed that it regulated apoptotic proteins (Bax and cleaved caspases 8 and 9) and autophagyrelated proteins (Beclin-1 and LC3B) but reduced the antiapoptosis proteins (Bcl-2; Figure 7B and C). Moreover, it was observed that CD31, a biomarker of tumor micro-vessels, 30 was downregulated in the tumor tissues after treatment ( Figure 7D), indicating an antiangiogenesis effect. The ICD effect was significantly induced by the LF-lipo treatment, as reflected by the increased CRT expression in the tumor sections ( Figure 7E). ...
Colorectal cancer (CRC) therapy is a big challenge, and seeking an effective and safe drug is a pressing clinical need. Gambogic acid is a potent antineoplastic agent without the drawback of bone marrow suppression. To improve its druggability (e.g., poor water solubility and tumor delivery), a lactoferrin-modified gambogic acid liposomal delivery system (LF-lipo) was developed to enhance the treatment efficacy of CRC. The LF-lipo can specifically bind LRP-1 expressed on colorectal cancer cells to enhance drug delivery to the tumor cells and yield enhanced therapeutic efficacy. The LF-lipo promoted tumor cell apoptosis and autophagy, reduced reactive oxygen species (ROS) levels in tumor cells, and inhibited angiogenesis; moreover, it could also repolarize tumor-associated macrophages from the M2 to M1 phenotype and induce ICD to activate T cells, exhibiting the capability of remodeling the tumor immune microenvironment. The liposomal formulation yielded an efficient and safe treatment outcome and has potential for clinical translation.
... We expected that assessing IMVD of STS at the time of diagnosis could give prognostic information for clinicians. In fact, angiogenic CD31 expression was correlated with chemotherapy resistance in sarcomas previously [53]. CD31 was also indicated to play a significant role in immune cell adhesion and integrin activation [53]. ...
... In fact, angiogenic CD31 expression was correlated with chemotherapy resistance in sarcomas previously [53]. CD31 was also indicated to play a significant role in immune cell adhesion and integrin activation [53]. At the same time, CD34-an endothelial cell marker-is also considered as one of the main activators of angiogenesis in sarcomas, including sarcoma recurrence [54]. ...
Background:
Marginally resectable and unresectable soft tissue sarcomas (STS) remain a therapy challenge due to the lack of highly active treatment. The aim of the study was to identify a biomarker to predict the pathological response (PR) to preplanned treatment of these STSs.
Methods:
In the phase II clinical trial (NCT03651375), locally advanced STS patients received preoperative treatment with a combination of doxorubicin-ifosfamide chemotherapy and 5 × 5 Gy radiotherapy. PR to the treatment was classified using the European Organization for Research and Treatment of Cancer-Soft Tissue and Bone Sarcoma Group recommendations. We have chosen HIF-1α, CD163, CD68, CD34, CD105, and γH2AFX proteins, rendering different biological phenomena, for biomarker study.
Results:
Nineteen patients were enrolled and in four cases a good PR was reported. The high expression of HIF-1α before surgery showed a negative correlation with PR, which means a poor response to therapy. Furthermore, the samples after surgery had decreased expression of HIF-1α, which confirmed the correlation with PR. However, high expression of γH2AFX positively correlated with PR, which provides better PR. The high number of positive-staining TAMs and the high IMVD did not correlate with PR.
Conclusions:
HIF1α and γH2AFX could be potential biomarkers for PR prediction after neoadjuvant treatment in STS.
... Maharani et al., 2018 [21] showed that pleomorphic visceral HSA cells from 17 dogs had decreased expression of CD 31 and vWF, and others have reported rare canine HSAs with loss of CD 31 expression [16,17,20] but expression of CD 31 has not been previously correlated with survival times. Venkataramani et al., 2018 [33] assessed CD 31 expression within tissue cultures derived from angiosarcomas, the human counterpart to animal hemangiosarcomas. They found that tumors expressing CD 31 LOW patterns represented prognostically worse tumors when compared with tumors expressing CD 31 HIGH . ...
... Therefore, maintenance or over-expression, as well as reduction or loss of cellular receptor expression such as CD 31, may both impart different growth advantages to neoplastic endothelial cells, but further studies are needed to determine their prognostic usefulness. In addition, correlating the success rate that chemotherapeutic agents extend the survival times of dogs with splenic HSA with tumor expression of CD 31 would be of interest, as CD 31 LOW endothelial cells in human angiosarcomas has been linked to resistance to various chemotherapeutic agents, including doxorubicin, which is used in the treatment of canine HSA [33]. ...
Canine hemangiosarcoma is a common, highly fatal tumor of older dogs, and predictors of survivability may provide clinically useful information. The objectives of this case series were to determine if a previously published tumor histological grading scheme, the level of tumor cellular atypia, clinical staging, or the level of CD 31 expression were useful for predicting the survival time in dogs with splenic hemangiosarcoma. Canine splenic hemangiosarcomas from 16 dogs were histologically graded, clinically staged, and assessed for CD 31 expression. Medical records were reviewed, the date of death was obtained, and survival data were analyzed statistically. Histopathological grading and clinical staging of canine splenic hemangiosarcomas, and the expression of CD 31 by the tumor cells were not significantly associated with the median survival time of the dogs in this study. However, strong expression of CD 31 by canine splenic hemangiosarcoma tumor cells was observed in dogs with short survival times, which warrants further studies to evaluate the potential prognostic value of CD 31 expression for the survival of dogs with splenic hemangiosarcoma.
... They are classified as a group of soft tissue sarcomas that are derived from lymphatic or vascular endothelial cells and are associated with a very poor prognosis [21,22]. Different versions of the CAM model have previously been used to successfully graft and culture angiosarcoma cell lines and test potential treatment agents [23][24][25]. ...
(1) Background: angiogenesis plays an important role in the growth and metastasis of tumors. We established the CAM assay application, an image analysis software of the IKOSA platform by KML Vision, for the quantification of blood vessels with the in ovo chorioallantoic membrane (CAM) model. We added this proprietary deep learning algorithm to the already established laser speckle contrast imaging (LSCI). (2) Methods: angiosarcoma cell line tumors were grafted onto the CAM. Angiogenesis was measured at the beginning and at the end of tumor growth with both measurement methods. The CAM assay application was trained to enable the recognition of in ovo CAM vessels. Histological stains of the tissue were performed and gluconate, an anti-angiogenic substance, was applied to the tumors. (3) Results: the angiosarcoma cells formed tumors on the CAM that appeared to stay vital and proliferated. An increase in perfusion was observed using both methods. The CAM assay application was successfully established in the in ovo CAM model and anti-angiogenic effects of gluconate were observed. (4) Conclusions: the CAM assay application appears to be a useful method for the quantification of angiogenesis in the CAM model and gluconate could be a potential treatment of angiosarcomas. Both aspects should be evaluated in further research.
... Our findings on reduced expression of CD31 in alcoholics (Table 2; Figure 2) might indicate the increased vascular stress in the microvascular bed. Furthermore, in a study in human angiosarcoma samples, Venkataramani et al. demonstrated that the downregulation of CD31 expression led to loss of endothelial tube formation and an increased induction of antioxidative enzymes [70]. In addition, it has been shown that in PD patients, apart from a decline in the vascular density, there is a morphological transformation in the vessel structure. ...
The blood–brain barrier (BBB) represents a highly specialized interface that acts as the first line of defense against toxins. Herein, we investigated the structural and ultrastructural changes in the basement membrane (BM), which is responsible for maintaining the integrity of the BBB, in the context of chronic alcoholism. Human post-mortem tissues from the Substantia Nigra (SN) region were obtained from 44 individuals, then grouped into controls, age-matched alcoholics, and non-age-matched alcoholics and assessed using light and electron microscopy. We found significantly less CD31+ vessels in alcoholic groups compared to controls in both gray and white matter samples. Alcoholics showed increased expression levels of collagen-IV, laminin-111, and fibronectin, which were coupled with a loss of BM integrity in comparison with controls. The BM of the gray matter was found to be more disintegrated than the white matter in alcoholics, as demonstrated by the expression of both collagen-IV and laminin-111, thereby indicating a breakdown in the BM’s structural composition. Furthermore, we observed that the expression of fibronectin was upregulated in the BM of the white matter vasculature in both alcoholic groups compared to controls. Taken together, our findings highlight some sort of aggregation or clumping of BM proteins that occurs in response to chronic alcohol consumption.
... We next wanted to test if the increase in nuclear YAP ratio in sparsely-plated hiCMs was correlated with higher YAP target gene expression. Thus, we curated a list of YAP target genes from the published literature [30][31][32] . We then compared the expression of these genes between densely-plated and sparsely-plated hiCMs. ...
... We turned to an MST1/2 inhibitor, XMU-MP-1, and a bioactive lipid that activates YAP, sphingosine-1-phosphate (S1P) 23,33,34 (Fig. 3A). Both of these compounds have been shown to upregulate YAP target genes in human embryonic stem cells, mouse liver cells and human hepatoma cells 30,33 . Furthermore, S1P increases Ki67 in densely plated hiCMs but not cell count when combined with lysophosphatidic acid 23 . ...
Inducing cardiac myocytes to proliferate is considered a potential therapy to target heart disease, however, modulating cardiac myocyte proliferation has proven to be a technical challenge. The Hippo pathway is a kinase signaling cascade that regulates cell proliferation during the growth of the heart. Inhibition of the Hippo pathway increases the activation of the transcription factors YAP/TAZ, which translocate to the nucleus and upregulate transcription of pro-proliferative genes. The Hippo pathway regulates the proliferation of cancer cells, pluripotent stem cells, and epithelial cells through a cell–cell contact-dependent manner, however, it is unclear if cell density-dependent cell proliferation is a consistent feature in cardiac myocytes. Here, we used cultured human iPSC-derived cardiac myocytes (hiCMs) as a model system to investigate this concept. hiCMs have a comparable transcriptome to the immature cardiac myocytes that proliferate during heart development in vivo. Our data indicate that a dense syncytium of hiCMs can regain cell cycle activity and YAP expression and activity when plated sparsely or when density is reduced through wounding. We found that combining two small molecules, XMU-MP-1 and S1P, increased YAP activity and further enhanced proliferation of low-density hiCMs. Importantly, these compounds had no effect on hiCMs within a dense syncytium. These data add to a growing body of literature that link Hippo pathway regulation with cardiac myocyte proliferation and demonstrate that regulation is restricted to cells with reduced contact inhibition.
... Moreover, there are no regular effective chemotherapy regimens. There is some promising data in exploring the potential role of the Hippo signaling pathway as a biological treatment of HA, the blockage of the vascular endothelial growth factor (VEGF) pathway, and even the use of beta-blockers [1,4,15,16]. ...
... Hopefully, the potential role of the Hippo signaling pathway, responsible for the regulation of cell proliferation and apoptosis, was explored in the biological treatment of angiosarcomas showing some promising results [4,15]. Yes-associated protein (YAP), an oncogene involved in the mentioned pathway, and CD31 regulate endothelial cell function and redox status via YAP [38]. ...
... Yes-associated protein (YAP), an oncogene involved in the mentioned pathway, and CD31 regulate endothelial cell function and redox status via YAP [38]. Angiosarcoma cells were subclassified based on the phenotypical expression, and the results showed that CD31 low was more common in angiosarcoma cases than CD31 high , and it was associated with YAP increase, making the tumor more chemoresistant to different agents, for example, doxorubicin [4,15]. Venkataramani et al. demonstrated in vitro that pazopanib, an effective YAP inhibitor, was effective when used with doxorubicin and resulted in resensitization of CD31 low to chemotherapy [15]. ...
Background:
Hepatic angiosarcoma is an uncommon, malignant, primary liver tumor, comprising 2% of liver cancers and accounting for < 1% of all sarcomas. Patients usually present with nonspecific symptoms, such as fatigue, weight loss, right upper quadrant pain, anemia, which leads to late diagnosis of an advanced stage tumor. The median life expectancy after the diagnosis of hepatic angiosarcoma is about 6 months, with only 3% of patients surviving more than 2 years. Liver failure and hemoperitoneum are the leading causes of death in patients with liver angiosarcoma. In rarer cases, it might cause paraneoplastic syndromes such as disseminated intravascular coagulopathy. The treatment of angiosarcomas is complicated as there are no established and effective treatment guidelines due to the tumor's low frequency and aggressive nature.
Case summary:
We present the case of a 68-year old woman who was admitted to the hospital due to fatigue and severe anemia (hemoglobin 65 g/l). Laboratory results also revealed high-grade thrombocytopenia (8 × 109/l). The abdominal ultrasound and computed tomography scan showed multiple lesions throughout the liver, spleen and kidneys. After the histological examination of the liver biopsy, the patient was diagnosed with hepatic angiosarcoma. The treatment with first-line chemotherapy (doxorubicin) was initiated despite ongoing paraneoplastic syndrome - disseminative intravascular coagulopathy. However, the disease was terminal, and the patient died 2 months since diagnosed.
Conclusions:
Hepatic angiosarcoma is a rare and terminal tumor. Therefore, knowledge about its manifestations and effective treatment methods is lacking. Disseminative intravascular coagulopathy is a unique clinical characteristic of angiosarcoma seen in a subset of patients.
... 29,30 In contrast, loss of CD31 expression in angiosarcomas is a sign of increased tumorigenesis and chemoresistance. 31 Furthermore, CTC clusters have been found to be more metastatic than single CTCs. 32 Taken together, these data indicate that CSV + CTCs may give prognostic information when examined beyond enumeration. ...
Background
Despite advances in care, the 5 year overall survival for patients with relapsed and or metastatic sarcoma remains as low as < 35%. Currently, there are no biomarkers available to assess disease status in patients with sarcomas and as such, disease surveillance remains reliant on serial imaging which increases the risk of secondary malignancies and heightens patient anxiety.
Methods
Here, for the first time reported in the literature, we have enumerated the cell surface vimentin (CSV+) CTCs in the blood of 92 sarcoma pediatric and adolescent and young adult (AYA) patients as a possible marker of disease.
Results
We constructed a ROC with an AUC of 0.831 resulting in a sensitivity of 85.3% and a specificity of 75%. Additionally, patients who were deemed to be CSV + CTC positive were found to have a worse overall survival compared to those who were CSV + CTC negative. We additionally found the use of available molecular testing increased the accuracy of our diagnostic and prognostic tests.
Conclusions
Our findings indicate that CSV + CTCs have both diagnostic and prognostic value and can possibly serve as a measure of disease burden.
... We also observed a significantly higher expression of surface marker CD31 (PECAM-1, platelet endothelial cell adhesion molecule-1) in the DPSC-12M. This increase can be explained by the action of free oxygen radicals [37] or in immature dental pulp stem cells [38]. It has been demonstrated that free oxygen radicals are also formed during the cryopreservation process [39]. ...
Dental pulp stem cells (DPSCs) are a type of easily accessible adult mesenchymal stem cell. Due to their ease of access, DPSCs show great promise in regenerative medicine. However, the tooth extractions from which DPSCs can be obtained are usually performed at a period of life when donors would have no therapeutic need of them. For this reason, it is imperative that successful stem cell storage techniques are employed so that these cells remain viable for future use. Any such techniques must result in high post-thaw stem cell recovery without compromising stemness, proliferation, or multipotency. Uncontrolled-rate freezing is not a technically or financially demanding technique compared to expensive and laborious controlled-rate freezing techniques. This study was aimed at observing the effect of uncontrolled-rate freezing on DPSCs stored for 6 and 12 months. Dimethyl sulfoxide at a concentration of 10% was used as a cryoprotective agent. Various features such as shape, proliferation capacity, phenotype, and multipotency were studied after DPSC thawing. The DPSCs did not compromise their stemness, viability, proliferation, or differentiating capabilities, even after one year of cryopreservation at -80 °C. After thawing, they retained their stemness markers and low-level expression of hematopoietic markers. We observed a size reduction in recovery DPSCs after one year of storage. This observation indicates that DPSCs can be successfully used in potential clinical applications, even after a year of uncontrolled cryopreservation.
