Pub Date : 2024-07-01Epub Date: 2024-09-10DOI: 10.1002/pgr2.70001
Mehmet Emre Erkanli, Ted Keunsil Kang, Thorsten Kirsch, Eva A Turley, Jin Ryoun Kim, Mary K Cowman
Peptides that increase pro-reparative responses to injury and disease by modulating the functional organization of hyaluronan (HA) with its cell surface binding proteins (e.g., soluble receptor for hyaluronan-mediated motility [RHAMM] and integral membrane CD44) have potential therapeutic value. The binding of RHAMM to HA is an attractive target, since RHAMM is normally absent or expressed at low levels in homeostatic conditions, but its expression is significantly elevated in the extracellular matrix during tissue stress, response-to-injury, and in cancers and inflammation-based diseases. The HA-binding site in RHAMM contains two closely spaced sequences of clustered basic amino acids, in an alpha-helical conformation. In the present communication, we test whether an alpha-helical conformation is required for effective peptide binding to HA, and competitive disruption of HA-RHAMM interaction. The HA-binding RHAMM-competitive peptide P15-1, identified using the unbiased approach of phage display, was examined using circular dichroism spectroscopy and the conformation-predictive AI-based AlphaFold2 algorithm. Unlike the HA-binding site in RHAMM, peptide P15-1 was found to adopt irregular conformations in solution rather than alpha helices. Instead, our structural analysis suggests that the primary determinant of peptide-HA binding is associated with a specific clustering and spacing pattern of basic amino acids, allowing favorable electrostatic interaction with carboxylate groups on HA.
通过调节透明质酸（HA）与其细胞表面结合蛋白（如透明质酸介导的可溶性受体[RHAMM]和完整膜CD44）的功能组织来增加对损伤和疾病的促复原反应的肽具有潜在的治疗价值。RHAMM 与 HA 的结合是一个极具吸引力的靶点，因为 RHAMM 通常不存在或在体内平衡状态下表达量较低，但在组织应激、损伤反应、癌症和炎症性疾病中，其在细胞外基质中的表达量会显著升高。RHAMM 中的 HA 结合位点包含两个紧密间隔的基本氨基酸序列，呈α-螺旋构象。在本通讯中，我们测试了α-螺旋构象是否是肽与 HA 有效结合以及竞争性破坏 HA-RHAMM 相互作用所必需的。我们使用圆二色性光谱和基于构象预测的人工智能 AlphaFold2 算法研究了使用噬菌体展示的无偏方法鉴定出的与 HA 结合的 RHAMM 竞争性多肽 P15-1。与 RHAMM 中的 HA 结合位点不同，肽 P15-1 在溶液中采用的是不规则构象，而不是α螺旋。相反，我们的结构分析表明，多肽与 HA 结合的主要决定因素与碱性氨基酸的特定聚类和间距模式有关，这种聚类和间距模式有利于与 HA 上的羧基发生静电相互作用。
{"title":"The spatial separation of basic amino acids is similar in RHAMM and hyaluronan binding peptide P15-1 despite different sequences and conformations.","authors":"Mehmet Emre Erkanli, Ted Keunsil Kang, Thorsten Kirsch, Eva A Turley, Jin Ryoun Kim, Mary K Cowman","doi":"10.1002/pgr2.70001","DOIUrl":"https://doi.org/10.1002/pgr2.70001","url":null,"abstract":"<p><p>Peptides that increase pro-reparative responses to injury and disease by modulating the functional organization of hyaluronan (HA) with its cell surface binding proteins (e.g., soluble receptor for hyaluronan-mediated motility [RHAMM] and integral membrane CD44) have potential therapeutic value. The binding of RHAMM to HA is an attractive target, since RHAMM is normally absent or expressed at low levels in homeostatic conditions, but its expression is significantly elevated in the extracellular matrix during tissue stress, response-to-injury, and in cancers and inflammation-based diseases. The HA-binding site in RHAMM contains two closely spaced sequences of clustered basic amino acids, in an alpha-helical conformation. In the present communication, we test whether an alpha-helical conformation is required for effective peptide binding to HA, and competitive disruption of HA-RHAMM interaction. The HA-binding RHAMM-competitive peptide P15-1, identified using the unbiased approach of phage display, was examined using circular dichroism spectroscopy and the conformation-predictive AI-based AlphaFold2 algorithm. Unlike the HA-binding site in RHAMM, peptide P15-1 was found to adopt irregular conformations in solution rather than alpha helices. Instead, our structural analysis suggests that the primary determinant of peptide-HA binding is associated with a specific clustering and spacing pattern of basic amino acids, allowing favorable electrostatic interaction with carboxylate groups on HA.</p>","PeriodicalId":74585,"journal":{"name":"Proteoglycan research","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11404675/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142302908","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-07-01Epub Date: 2024-06-29DOI: 10.1002/pgr2.22
Maurizio Mongiat, Gabriel Pascal, Evelina Poletto, Davion M Williams, Renato V Iozzo
Anti-angiogenic therapy is an established method for the treatment of several cancers and vascular-related diseases. Most of the agents employed target the vascular endothelial growth factor A, the major cytokine stimulating angiogenesis. However, the efficacy of these treatments is limited by the onset of drug resistance. Therefore, it is of fundamental importance to better understand the mechanisms that regulate angiogenesis and the microenvironmental cues that play significant role and influence patient treatment and outcome. In this context, here we review the importance of the three basement membrane heparan sulfate proteoglycans (HSPGs), namely perlecan, agrin and collagen XVIII. These HSPGs are abundantly expressed in the vasculature and, due to their complex molecular architecture, they interact with multiple endothelial cell receptors, deeply affecting their function. Under normal conditions, these proteoglycans exert pro-angiogenic functions. However, in pathological conditions such as cancer and inflammation, extracellular matrix remodeling leads to the degradation of these large precursor molecules and the liberation of bioactive processed fragments displaying potent angiostatic activity. These unexpected functions have been demonstrated for the C-terminal fragments of perlecan and collagen XVIII, endorepellin and endostatin. These bioactive fragments can also induce autophagy in vascular endothelial cells which contributes to angiostasis. Overall, basement membrane proteoglycans deeply affect angiogenesis counterbalancing pro-angiogenic signals during tumor progression, and represent possible means to develop new prognostic biomarkers and novel therapeutic approaches for the treatment of solid tumors.
{"title":"Proteoglycans of basement membranes: Crucial controllers of angiogenesis, neurogenesis, and autophagy.","authors":"Maurizio Mongiat, Gabriel Pascal, Evelina Poletto, Davion M Williams, Renato V Iozzo","doi":"10.1002/pgr2.22","DOIUrl":"10.1002/pgr2.22","url":null,"abstract":"<p><p>Anti-angiogenic therapy is an established method for the treatment of several cancers and vascular-related diseases. Most of the agents employed target the vascular endothelial growth factor A, the major cytokine stimulating angiogenesis. However, the efficacy of these treatments is limited by the onset of drug resistance. Therefore, it is of fundamental importance to better understand the mechanisms that regulate angiogenesis and the microenvironmental cues that play significant role and influence patient treatment and outcome. In this context, here we review the importance of the three basement membrane heparan sulfate proteoglycans (HSPGs), namely perlecan, agrin and collagen XVIII. These HSPGs are abundantly expressed in the vasculature and, due to their complex molecular architecture, they interact with multiple endothelial cell receptors, deeply affecting their function. Under normal conditions, these proteoglycans exert pro-angiogenic functions. However, in pathological conditions such as cancer and inflammation, extracellular matrix remodeling leads to the degradation of these large precursor molecules and the liberation of bioactive processed fragments displaying potent angiostatic activity. These unexpected functions have been demonstrated for the C-terminal fragments of perlecan and collagen XVIII, endorepellin and endostatin. These bioactive fragments can also induce autophagy in vascular endothelial cells which contributes to angiostasis. Overall, basement membrane proteoglycans deeply affect angiogenesis counterbalancing pro-angiogenic signals during tumor progression, and represent possible means to develop new prognostic biomarkers and novel therapeutic approaches for the treatment of solid tumors.</p>","PeriodicalId":74585,"journal":{"name":"Proteoglycan research","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11340296/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142057454","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Yingying Xu, Yi Liu, Ruyi Zou, Xunyi Yuan, Lin Wei, Yong Qin, Xu Wang, Yunxue Zhao, Zhenqiu Yang, Wenshuang Wang, Fuchuan Li
The synthesis of glycosaminoglycans (GAGs) in vivo occurs with high spatiotemporal specificity, and any aberrant expression of GAGs is closely related to the occurrence of diseases. In terms of tumorigenesis, the abnormally expressed GAGs have become a potential target for the diagnosis and therapy of tumors. As previously reported, VAR2HP, a protein probe that recognizes the unique heparin (Hep)‐like epitopes, interacts with a decasaccharide structure containing at least three HexA2S(1‐4)GlcNS6S disaccharides. Its recognition epitopes are overexpressed in various tumor cells and appear promising as target molecules of multiple tumors. Herein, we found that VAR2HP used as an antineoplastic carrier could promote drug enrichment in tumor sites by targeting the specific Hep‐like epitopes on tumor cells, thereby reducing the damage to normal cells in vitro and in vivo. Moreover, VAR2HP acting on cells alone could inhibit cell proliferation, indicating that VAR2HP as a drug carrier has a dual effect of antitumor activity. Additionally, we observed that VAR2HP significantly stained cancer tissues more strongly than neighboring nonmalignant tissues. The staining was competitively inhibited by added exogenous Hep, indicating that the Hep‐like epitopes recognized by VAR2HP were a potential target molecule in tumor diagnosis. Moreover, the VAR2HP‐bound Hep‐like epitopes were found to be overexpressed in the sera of patients with hepatocellular carcinoma (HCC) but not in normal persons and patients with cirrhosis. Taken together, this study shows that VAR2HP and its heparin‐like epitopes have great potential in targeted therapy of tumors and HCC diagnosis.
体内糖胺聚糖（GAGs）的合成具有高度的时空特异性，GAGs的异常表达与疾病的发生密切相关。在肿瘤发生方面，异常表达的 GAGs 已成为肿瘤诊断和治疗的潜在靶点。正如之前所报道的，VAR2HP 是一种能识别独特肝素（Hep）样表位的蛋白探针，它与至少含有三个 HexA2S(1-4)GlcNS6S 二糖的十糖结构相互作用。其识别表位在多种肿瘤细胞中过度表达，有望成为多种肿瘤的靶分子。在此，我们发现，VAR2HP 作为抗肿瘤载体，可通过靶向肿瘤细胞上的特异性 Hep 类表位，促进药物在肿瘤部位的富集，从而减少体外和体内对正常细胞的损伤。此外，VAR2HP 单独作用于细胞可抑制细胞增殖，表明 VAR2HP 作为药物载体具有抗肿瘤活性的双重作用。此外，我们还观察到 VAR2HP 对癌症组织的染色明显强于邻近的非恶性组织。添加外源 Hep 可竞争性抑制该染色，这表明 VAR2HP 识别的 Hep 类表位是肿瘤诊断的潜在靶分子。此外，还发现 VAR2HP 结合的 Hep 类表位在肝细胞癌（HCC）患者血清中过度表达，而在正常人和肝硬化患者血清中则没有。综上所述，本研究表明 VAR2HP 及其肝素样表位在肿瘤靶向治疗和 HCC 诊断方面具有巨大潜力。
{"title":"VAR2HP recognizing heparin‐like epitopes in targeted therapy and diagnosis of tumors","authors":"Yingying Xu, Yi Liu, Ruyi Zou, Xunyi Yuan, Lin Wei, Yong Qin, Xu Wang, Yunxue Zhao, Zhenqiu Yang, Wenshuang Wang, Fuchuan Li","doi":"10.1002/pgr2.16","DOIUrl":"https://doi.org/10.1002/pgr2.16","url":null,"abstract":"The synthesis of glycosaminoglycans (GAGs) in vivo occurs with high spatiotemporal specificity, and any aberrant expression of GAGs is closely related to the occurrence of diseases. In terms of tumorigenesis, the abnormally expressed GAGs have become a potential target for the diagnosis and therapy of tumors. As previously reported, VAR2HP, a protein probe that recognizes the unique heparin (Hep)‐like epitopes, interacts with a decasaccharide structure containing at least three HexA2S(1‐4)GlcNS6S disaccharides. Its recognition epitopes are overexpressed in various tumor cells and appear promising as target molecules of multiple tumors. Herein, we found that VAR2HP used as an antineoplastic carrier could promote drug enrichment in tumor sites by targeting the specific Hep‐like epitopes on tumor cells, thereby reducing the damage to normal cells in vitro and in vivo. Moreover, VAR2HP acting on cells alone could inhibit cell proliferation, indicating that VAR2HP as a drug carrier has a dual effect of antitumor activity. Additionally, we observed that VAR2HP significantly stained cancer tissues more strongly than neighboring nonmalignant tissues. The staining was competitively inhibited by added exogenous Hep, indicating that the Hep‐like epitopes recognized by VAR2HP were a potential target molecule in tumor diagnosis. Moreover, the VAR2HP‐bound Hep‐like epitopes were found to be overexpressed in the sera of patients with hepatocellular carcinoma (HCC) but not in normal persons and patients with cirrhosis. Taken together, this study shows that VAR2HP and its heparin‐like epitopes have great potential in targeted therapy of tumors and HCC diagnosis.","PeriodicalId":74585,"journal":{"name":"Proteoglycan research","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140520843","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Paraskevi Ioannou, Kyriaki Tzaferi, Christos Koutsakis, Z. Piperigkou, Nikos K. Karamanos
Extracellular matrix (ECM) and its dynamic remodeling contribute to the progression of breast cancer, the most prevailing cancer type in women. Glypicans (GPCs) function as cell co‐receptors by facilitating the formation of ligand–receptor complexes. An important regulator in the context of breast cancer progression is the JAK/STAT signaling pathway that oversees the expression of genes associated with cancer cell characteristics. Epidermal growth factor receptor (EGFR) is a pivotal player in this process. The aim of this study is to examine the effect of the EGFR and JAK/STAT signaling pathways on GPCs expression in breast cancer cells with different estrogen receptor (ER) status, depicting different breast cancer subtypes. To this end, the ERα‐positive MCF‐7, and the ERβ‐positive MDA‐MB‐231 breast cancer cell lines were evaluated in terms of the impact of downstream inhibition of both pathways on the functional properties as well as the expression of GPCs 1‐6 genes. Notably, the downstream inhibition of both EGFR and JAK/STAT cascades mitigate cell proliferation and migration, while increasing cell adhesion on collagen type I in an ER‐independent manner. However, the inhibition exhibited a cell‐line‐dependent effect on GPC expression, as in MCF‐7 cells GPCs expression is mostly downregulated excepting GPC‐4 and GPC‐5. Conversely, in MDA‐MB‐231 cells, EGFR and JAK/STAT activation is essential for maintaining GPCs at low levels. Additionally, STRING analysis identified the small leucine‐rich PG decorin as a putative link between all GPCs and EGFR. Subsequently, a deeper understanding on the effect of EGFR and JAK/STAT signaling may shed light into the role and interplay between GPCs and decorin in breast cancer progression, thus contributing to novel therapeutic solutions.
{"title":"Targeting glypicans through EGFR and JAK/STAT signaling axes drives breast cancer progression","authors":"Paraskevi Ioannou, Kyriaki Tzaferi, Christos Koutsakis, Z. Piperigkou, Nikos K. Karamanos","doi":"10.1002/pgr2.18","DOIUrl":"https://doi.org/10.1002/pgr2.18","url":null,"abstract":"Extracellular matrix (ECM) and its dynamic remodeling contribute to the progression of breast cancer, the most prevailing cancer type in women. Glypicans (GPCs) function as cell co‐receptors by facilitating the formation of ligand–receptor complexes. An important regulator in the context of breast cancer progression is the JAK/STAT signaling pathway that oversees the expression of genes associated with cancer cell characteristics. Epidermal growth factor receptor (EGFR) is a pivotal player in this process. The aim of this study is to examine the effect of the EGFR and JAK/STAT signaling pathways on GPCs expression in breast cancer cells with different estrogen receptor (ER) status, depicting different breast cancer subtypes. To this end, the ERα‐positive MCF‐7, and the ERβ‐positive MDA‐MB‐231 breast cancer cell lines were evaluated in terms of the impact of downstream inhibition of both pathways on the functional properties as well as the expression of GPCs 1‐6 genes. Notably, the downstream inhibition of both EGFR and JAK/STAT cascades mitigate cell proliferation and migration, while increasing cell adhesion on collagen type I in an ER‐independent manner. However, the inhibition exhibited a cell‐line‐dependent effect on GPC expression, as in MCF‐7 cells GPCs expression is mostly downregulated excepting GPC‐4 and GPC‐5. Conversely, in MDA‐MB‐231 cells, EGFR and JAK/STAT activation is essential for maintaining GPCs at low levels. Additionally, STRING analysis identified the small leucine‐rich PG decorin as a putative link between all GPCs and EGFR. Subsequently, a deeper understanding on the effect of EGFR and JAK/STAT signaling may shed light into the role and interplay between GPCs and decorin in breast cancer progression, thus contributing to novel therapeutic solutions.","PeriodicalId":74585,"journal":{"name":"Proteoglycan research","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140524082","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
V. Masola, G. Marrone, Carola Condoluci, Marco Franchi, Leonardo Stella, M. Biolato, Luca Miele, Giovanni Gambaro, Claudia Dal Vecchio, M. Onisto
Together with the ACE2 protein, heparan sulfate present at the level of the glycocalyx in the lung epithelia is considered a cellular “co‐receptor” for the viral spike protein that allows severe acute respiratory syndrome coronavirus 2 (SARS‐CoV) to infect cells. An increase in the amount and activity of heparanase‐1 (HPSE), the only enzyme capable of degrading the heparan sulfate (HS) chains of the glycocalyx and of the extracellular matrix, has been described in the plasma of patients affected by coronavirus disease 2019 (Covid‐19). Furthermore, the activity of matrix metalloproteases, or MMPs, has been related to matrix degradation, oxidative stress, and inflammation in Covid‐19 patients. In this study, we enrolled 26 Covid‐19 patients and 15 controls with diagnosis of non‐SARS‐CoV‐2‐related pneumonia. We evaluated the expression and activity of HPSE and the expression of MMPs in their serum together with other clinical markers of disease and inflammation. Results proved that HPSE expression and activity serum levels were significantly increased, whereas MMP2 and 9 were decreased in Covid‐19 versus non‐Covid‐19 pneumonia patients. In addition, IL‐6 levels were higher, whereas platelet and white blood cells were lower in Covid‐19 with respect to non‐Covid‐19 pneumonia. Moreover, MMP9 but not HPSE (expression and activity) levels were increased in Covid‐19 pneumonia patients with ongoing lung alteration over time. In summary, the present findings indicate that HPSE and MMPs are differentially regulated in Covid‐19 and non‐Covid‐19 pneumonia.
{"title":"Heparanase‐1 and MMPs in Covid‐19 and non‐Covid‐19 pneumonia","authors":"V. Masola, G. Marrone, Carola Condoluci, Marco Franchi, Leonardo Stella, M. Biolato, Luca Miele, Giovanni Gambaro, Claudia Dal Vecchio, M. Onisto","doi":"10.1002/pgr2.14","DOIUrl":"https://doi.org/10.1002/pgr2.14","url":null,"abstract":"Together with the ACE2 protein, heparan sulfate present at the level of the glycocalyx in the lung epithelia is considered a cellular “co‐receptor” for the viral spike protein that allows severe acute respiratory syndrome coronavirus 2 (SARS‐CoV) to infect cells. An increase in the amount and activity of heparanase‐1 (HPSE), the only enzyme capable of degrading the heparan sulfate (HS) chains of the glycocalyx and of the extracellular matrix, has been described in the plasma of patients affected by coronavirus disease 2019 (Covid‐19). Furthermore, the activity of matrix metalloproteases, or MMPs, has been related to matrix degradation, oxidative stress, and inflammation in Covid‐19 patients. In this study, we enrolled 26 Covid‐19 patients and 15 controls with diagnosis of non‐SARS‐CoV‐2‐related pneumonia. We evaluated the expression and activity of HPSE and the expression of MMPs in their serum together with other clinical markers of disease and inflammation. Results proved that HPSE expression and activity serum levels were significantly increased, whereas MMP2 and 9 were decreased in Covid‐19 versus non‐Covid‐19 pneumonia patients. In addition, IL‐6 levels were higher, whereas platelet and white blood cells were lower in Covid‐19 with respect to non‐Covid‐19 pneumonia. Moreover, MMP9 but not HPSE (expression and activity) levels were increased in Covid‐19 pneumonia patients with ongoing lung alteration over time. In summary, the present findings indicate that HPSE and MMPs are differentially regulated in Covid‐19 and non‐Covid‐19 pneumonia.","PeriodicalId":74585,"journal":{"name":"Proteoglycan research","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140523712","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
E. Nakato, Sarah Baker, A. Kinoshita-Toyoda, Collin Knudsen, Yi‐Si Lu, Masahiko Takemura, Hidenao Toyoda, H. Nakato
Heparan sulfate proteoglycans (HSPGs) serve as co‐receptors for growth factor signaling during development. It is well known that the level and patterns of sulfate groups of heparan sulfate (HS) chains, or HS fine structures, have a major impact on HSPG function. On the other hand, the physiological significance of other structural features of HS, including NS/NA domain organization, remains to be elucidated. A blueprint of the HS domain structures is mainly controlled by HS N‐deacetylase/N‐sulfotransferases (NDSTs). To analyze in vivo activities of differentially modified HS, we established two knock‐in (KI) Drosophila strains with the insertion of mouse Ndst1 (mNdst1) or Ndst2 (mNdst2) in the locus of sulfateless (sfl), the only Drosophila NDST. In these KI lines, mNDSTs are expressed from the sfl locus, in the level and patterns identical to the endogenous sfl gene. Thus, phenotypes of Ndst1 KI and Ndst2KI animals reflect the ability of HS structures made by these enzymes to rescue sfl mutation. Remarkably, we found that mNdst1 completely rescued the loss of sfl. mNdst2 showed a limited rescue ability, despite a higher level of HS sulfation compared to HS in mNdst1 KI. Our study suggests that independent of sulfation levels, additional HS structural features controlled by NDSTs play key roles during tissue patterning.
{"title":"In vivo activities of heparan sulfate differentially modified by NDSTs during development","authors":"E. Nakato, Sarah Baker, A. Kinoshita-Toyoda, Collin Knudsen, Yi‐Si Lu, Masahiko Takemura, Hidenao Toyoda, H. Nakato","doi":"10.1002/pgr2.17","DOIUrl":"https://doi.org/10.1002/pgr2.17","url":null,"abstract":"Heparan sulfate proteoglycans (HSPGs) serve as co‐receptors for growth factor signaling during development. It is well known that the level and patterns of sulfate groups of heparan sulfate (HS) chains, or HS fine structures, have a major impact on HSPG function. On the other hand, the physiological significance of other structural features of HS, including NS/NA domain organization, remains to be elucidated. A blueprint of the HS domain structures is mainly controlled by HS N‐deacetylase/N‐sulfotransferases (NDSTs). To analyze in vivo activities of differentially modified HS, we established two knock‐in (KI) Drosophila strains with the insertion of mouse Ndst1 (mNdst1) or Ndst2 (mNdst2) in the locus of sulfateless (sfl), the only Drosophila NDST. In these KI lines, mNDSTs are expressed from the sfl locus, in the level and patterns identical to the endogenous sfl gene. Thus, phenotypes of Ndst1 KI and Ndst2KI animals reflect the ability of HS structures made by these enzymes to rescue sfl mutation. Remarkably, we found that mNdst1 completely rescued the loss of sfl. mNdst2 showed a limited rescue ability, despite a higher level of HS sulfation compared to HS in mNdst1 KI. Our study suggests that independent of sulfation levels, additional HS structural features controlled by NDSTs play key roles during tissue patterning.","PeriodicalId":74585,"journal":{"name":"Proteoglycan research","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140524922","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Sofia Nizzolo, Emiliano Esposito, Ming‐Hong Ni, Laura Bertocchi, Giulio Bianchini, Nadia Freato, Serena Zanzoni, Marco Guerrini, Sabrina Bertini
Sodium hyaluronate (HA), a derivative of hyaluronan, is a natural and biocompatible polysaccharide that interacts with cluster of differentiation‐44 receptor to promote fine‐tuning of inflammation, fibrosis, and tissue remodeling. HA has a smaller molecular weight than hyaluronan and is overall more stable being less prone to oxidation. In this study, we report a novel lactose‐functionalized sodium hyaluronate, named HYLACH®. Functionalization with multiple β‐galactose residues facilitates its interaction with galectin‐3, a β‐galactose binding lectin implicated in various pathological processes including inflammation, host defense, and fibrosis, especially critical in idiopathic pulmonary fibrosis (IPF). Our strategy was to modify HA, to varying extents, at carboxyl sites with 1‐amino‐1‐deoxy‐lactitol, in the presence of 4‐(4,6‐dimethoxy‐1,3,5‐triazin‐2‐yl)‐4‐methyl morpholinium chloride in aqueous media. We characterized the chemical structure, molecular weight, and degree of substitution of HYLACH® using NMR spectroscopy and size exclusion chromatography. We further determined several key parameters including its stability toward enzymatic degradation and the binding affinity and conformational changes of galectin‐3 interaction with HYLACH®. Collectively, the generation of a novel functionalized HA with an ability to bind and suppress galectin‐3 function, in combination with safety and biocompatibility, offers the opportunity to test this compound in therapeutic trials of devastating fibrotic diseases such as IPF.
透明质酸钠（HA）是透明质酸的衍生物，是一种天然的生物相容性多糖，可与分化-44 受体群相互作用，促进炎症、纤维化和组织重塑的微调。与透明质酸相比，HA 的分子量更小，而且总体上更稳定，不易氧化。在这项研究中，我们报告了一种新型乳糖功能化透明质酸钠，名为 HYLACH®。多个β-半乳糖残基的功能化促进了透明质酸钠与半乳糖凝集素-3的相互作用，半乳糖凝集素-3是一种与炎症、宿主防御和纤维化等各种病理过程有关的β-半乳糖结合凝集素，在特发性肺纤维化（IPF）中尤为关键。我们的策略是在水介质中，在 4-(4,6-二甲氧基-1,3,5-三嗪-2-基)-4-甲基吗啉氯化物的存在下，用 1-氨基-1-脱氧-乳糖醇对 HA 的羧基位点进行不同程度的修饰。我们利用核磁共振光谱和尺寸排阻色谱法确定了 HYLACH® 的化学结构、分子量和取代度。我们还进一步确定了几个关键参数，包括其酶降解稳定性、结合亲和力以及 galectin-3 与 HYLACH® 相互作用的构象变化。总之，新型功能化 HA 的产生具有结合和抑制 galectin-3 功能的能力，同时兼具安全性和生物相容性，这为在 IPF 等破坏性纤维化疾病的治疗试验中测试这种化合物提供了机会。
{"title":"A novel biomimetic probe for galectin‐3 recognition: Chemical synthesis and structural characterization of a β‐galactose branched sodium hyaluronate","authors":"Sofia Nizzolo, Emiliano Esposito, Ming‐Hong Ni, Laura Bertocchi, Giulio Bianchini, Nadia Freato, Serena Zanzoni, Marco Guerrini, Sabrina Bertini","doi":"10.1002/pgr2.19","DOIUrl":"https://doi.org/10.1002/pgr2.19","url":null,"abstract":"Sodium hyaluronate (HA), a derivative of hyaluronan, is a natural and biocompatible polysaccharide that interacts with cluster of differentiation‐44 receptor to promote fine‐tuning of inflammation, fibrosis, and tissue remodeling. HA has a smaller molecular weight than hyaluronan and is overall more stable being less prone to oxidation. In this study, we report a novel lactose‐functionalized sodium hyaluronate, named HYLACH®. Functionalization with multiple β‐galactose residues facilitates its interaction with galectin‐3, a β‐galactose binding lectin implicated in various pathological processes including inflammation, host defense, and fibrosis, especially critical in idiopathic pulmonary fibrosis (IPF). Our strategy was to modify HA, to varying extents, at carboxyl sites with 1‐amino‐1‐deoxy‐lactitol, in the presence of 4‐(4,6‐dimethoxy‐1,3,5‐triazin‐2‐yl)‐4‐methyl morpholinium chloride in aqueous media. We characterized the chemical structure, molecular weight, and degree of substitution of HYLACH® using NMR spectroscopy and size exclusion chromatography. We further determined several key parameters including its stability toward enzymatic degradation and the binding affinity and conformational changes of galectin‐3 interaction with HYLACH®. Collectively, the generation of a novel functionalized HA with an ability to bind and suppress galectin‐3 function, in combination with safety and biocompatibility, offers the opportunity to test this compound in therapeutic trials of devastating fibrotic diseases such as IPF.","PeriodicalId":74585,"journal":{"name":"Proteoglycan research","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140515769","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
A proliferation‐inducing ligand (APRIL), a member of the tumor necrosis factor superfamily, affects the survival and proliferation of tumor cells. Understanding the mechanism of action of APRIL in tumor cells, including intracellular signaling, is important for its potential use in diagnostics and prognosis. It has been shown that APRIL‐induced tumor proliferation requires heparan sulfate (HS) proteoglycans to mediate the binding of APRIL to tumor cells. Here, we show that chondroitin sulfate (CS) proteoglycan mainly contributes to the APRIL‐stimulated proliferation of triple‐negative breast cancer BT‐549 cells. Knockout of chondroitin 4‐O‐sulfotransferase‐1 (C4ST‐1), a key CS biosynthetic enzyme, suppressed APRIL‐induced tumor proliferation, whereas deficiency of exostosin 1 (EXT1), a key HS biosynthetic enzyme, had only weak effects. Molecular interaction analyses using Biacore revealed that although CS did not directly bind to tumor growth through Ca2+ modulator interactor (TACI), it enhanced the binding of APRIL to the APRIL receptor, TACI. The small leucine‐rich proteoglycan, biglycan, plays a pivotal role in tumor growth and progression. Biglycan knockdown inhibited BT‐549 cell proliferation. These results suggest that CS synthesized by C4ST‐1 participates in APRIL signaling and modulates pathological events in tumors.
增殖诱导配体（APRIL）是肿瘤坏死因子超家族的成员之一，会影响肿瘤细胞的存活和增殖。了解 APRIL 在肿瘤细胞中的作用机制（包括细胞内信号传导）对其在诊断和预后中的潜在应用非常重要。研究表明，APRIL 诱导的肿瘤增殖需要硫酸肝素（HS）蛋白多糖介导 APRIL 与肿瘤细胞的结合。在这里，我们发现硫酸软骨素（CS）蛋白多糖主要促进了三阴性乳腺癌BT-549细胞在APRIL刺激下的增殖。敲除软骨素 4-O-磺基转移酶-1（C4ST-1）--一种关键的 CS 生物合成酶--抑制了 APRIL 诱导的肿瘤增殖，而缺乏外ostosin 1（EXT1）--一种关键的 HS 生物合成酶--只有微弱的影响。利用 Biacore 进行的分子相互作用分析表明，尽管 CS 没有通过 Ca2+ 调制剂相互作用因子（TACI）直接与肿瘤生长结合，但它增强了 APRIL 与 APRIL 受体 TACI 的结合。富含亮氨酸的小蛋白多糖（biglycan）在肿瘤生长和进展中起着关键作用。敲除 Biglycan 可抑制 BT-549 细胞的增殖。这些结果表明，C4ST-1合成的CS参与了APRIL信号转导，并调节肿瘤的病理事件。
{"title":"Chondroitin sulfate proteoglycan promotes APRIL‐induced tumor cell proliferation","authors":"S. Nadanaka, Toshiyasu Koike, Hiroshi Kitagawa","doi":"10.1002/pgr2.15","DOIUrl":"https://doi.org/10.1002/pgr2.15","url":null,"abstract":"A proliferation‐inducing ligand (APRIL), a member of the tumor necrosis factor superfamily, affects the survival and proliferation of tumor cells. Understanding the mechanism of action of APRIL in tumor cells, including intracellular signaling, is important for its potential use in diagnostics and prognosis. It has been shown that APRIL‐induced tumor proliferation requires heparan sulfate (HS) proteoglycans to mediate the binding of APRIL to tumor cells. Here, we show that chondroitin sulfate (CS) proteoglycan mainly contributes to the APRIL‐stimulated proliferation of triple‐negative breast cancer BT‐549 cells. Knockout of chondroitin 4‐O‐sulfotransferase‐1 (C4ST‐1), a key CS biosynthetic enzyme, suppressed APRIL‐induced tumor proliferation, whereas deficiency of exostosin 1 (EXT1), a key HS biosynthetic enzyme, had only weak effects. Molecular interaction analyses using Biacore revealed that although CS did not directly bind to tumor growth through Ca2+ modulator interactor (TACI), it enhanced the binding of APRIL to the APRIL receptor, TACI. The small leucine‐rich proteoglycan, biglycan, plays a pivotal role in tumor growth and progression. Biglycan knockdown inhibited BT‐549 cell proliferation. These results suggest that CS synthesized by C4ST‐1 participates in APRIL signaling and modulates pathological events in tumors.","PeriodicalId":74585,"journal":{"name":"Proteoglycan research","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139633520","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Versican is a large chondroitin sulfate/dermatan sulfate proteoglycan in the extracellular matrix and one of the aggrecan/lectican family. Whereas versican is constitutively expressed and serves as a structural macromolecule in some tissues, it is transiently expressed at high levels when the extracellular matrix dynamically changes. There, versican plays an important role in forming the provisional matrix, which is replaced with the “authentic” extracellular matrix, that is, the matrix as it should be. ADAMTS‐1, 4, 5, 9, 15, and 20 cleave versican core protein and are therefore named versicanases. These proteinases have been believed to play a critical role in versican turnover. A cleaved N‐terminal fragment harbors biological functions, and it is termed “versikine.” This review discusses recent advances in the research on the in vivo function of versican and versikine generated by versicanases.
{"title":"Versican and versikine: The dynamism of the extracellular matrix","authors":"Hideto Watanabe","doi":"10.1002/pgr2.13","DOIUrl":"https://doi.org/10.1002/pgr2.13","url":null,"abstract":"Versican is a large chondroitin sulfate/dermatan sulfate proteoglycan in the extracellular matrix and one of the aggrecan/lectican family. Whereas versican is constitutively expressed and serves as a structural macromolecule in some tissues, it is transiently expressed at high levels when the extracellular matrix dynamically changes. There, versican plays an important role in forming the provisional matrix, which is replaced with the “authentic” extracellular matrix, that is, the matrix as it should be. ADAMTS‐1, 4, 5, 9, 15, and 20 cleave versican core protein and are therefore named versicanases. These proteinases have been believed to play a critical role in versican turnover. A cleaved N‐terminal fragment harbors biological functions, and it is termed “versikine.” This review discusses recent advances in the research on the in vivo function of versican and versikine generated by versicanases.","PeriodicalId":74585,"journal":{"name":"Proteoglycan research","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139330868","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Abstract Biglycan is a member of the small leucine‐rich proteoglycan family. Dysregulation of biglycan leads to a broad range of clinical consequences, such as osteoclastogenesis, inflammation, cardiovascular disease, and cancer. Biglycan binding to toll‐like receptor (TLR)−2 or TLR‐4 on immune cells lead to infiltration of immune cells to mediate the inflammatory response. Additionally, the extracellular matrix‐secreted soluble biglycan functions as a danger‐associated molecular pattern molecule involved in the induction of inflammation and cancer. High expression of biglycan is demonstrated in tumor endothelial cells (TECs) of various cancers and correlates with metastatic potential and poor clinical outcomes. This comprehensive review addresses the role of biglycan in both tumor cells and tumor stromal cells, especially TECs, in regulating tumor angiogenesis, tumor growth, metastasis, and chemotherapy resistance.
{"title":"The functional network of biglycan: A new frontier in tumor progression","authors":"Li Yu, Nako Maishi, Aya Matsuda, Kyoko Hida","doi":"10.1002/pgr2.11","DOIUrl":"https://doi.org/10.1002/pgr2.11","url":null,"abstract":"Abstract Biglycan is a member of the small leucine‐rich proteoglycan family. Dysregulation of biglycan leads to a broad range of clinical consequences, such as osteoclastogenesis, inflammation, cardiovascular disease, and cancer. Biglycan binding to toll‐like receptor (TLR)−2 or TLR‐4 on immune cells lead to infiltration of immune cells to mediate the inflammatory response. Additionally, the extracellular matrix‐secreted soluble biglycan functions as a danger‐associated molecular pattern molecule involved in the induction of inflammation and cancer. High expression of biglycan is demonstrated in tumor endothelial cells (TECs) of various cancers and correlates with metastatic potential and poor clinical outcomes. This comprehensive review addresses the role of biglycan in both tumor cells and tumor stromal cells, especially TECs, in regulating tumor angiogenesis, tumor growth, metastasis, and chemotherapy resistance.","PeriodicalId":74585,"journal":{"name":"Proteoglycan research","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136260470","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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