bims-supasi 2023-04-23 papers

bims-supasi

Biomed News

on Sulfation pathways and signalling

Issue of 2023–04–23
seven papers selected by
Jonathan Wolf Mueller, University of Birmingham

Increased Cerebral Serum Amyloid A2 and Parameters of Oxidation in Arylsulfatase B (N-Acetylgalactosamine-4-Sulfatase)-Null Mice.
TFCP2 is a transcriptional regulator of heparan sulfate assembly and melanoma cell growth.
SARS-CoV-2 evolved variants optimize binding to cellular glycocalyx.
Novel (sulfated) thyroid hormone transporters in the solute carrier 22 family.
Acute changes in serum and skeletal muscle steroids in resistance-trained men.
The role of KS GAGs in the microstructure of CXL-treated corneal stroma; a transmission electron microscopy study.
Analyzing the effect of heparin on in vitro capacitation and spermatozoal RNA population in goats.

bioRxiv. 2023 Apr 03. pii: 2023.04.03.535377. [Epub ahead of print]

Increased Cerebral Serum Amyloid A2 and Parameters of Oxidation in Arylsulfatase B (N-Acetylgalactosamine-4-Sulfatase)-Null Mice.

Sumit Bhattacharyya, Joanne K Tobacman.

Introduction: Chondroitin sulfate and chondroitin sulfate proteoglycans have been associated with Alzheimer's Disease (AD), and the impact of modified chondroitin sulfates is being investigated in several animal and cell-based models of AD. Published reports have shown the role of accumulation of chondroitin 4-sulfate and decline in Arylsulfatase B (ARSB; B-acetylgalactosamine-4-sulfatase) in other pathology, including nerve injury, traumatic brain injury, and spinal cord injury. However, the impact of ARSB deficiency on AD pathobiology has not been reported, although changes in ARSB were associated with AD in two prior reports. The enzyme ARSB removes 4-sulfate groups from the non-reducing end of chondroitin 4-sulfate and dermatan sulfate and is required for their degradation. When ARSB activity declines, these sulfated glycosaminoglycans accumulate, as in the inherited disorder Mucopolysaccharidosis VI.
Methods: Reports about chondroitin sulfate, chondroitin sulfate proteoglycans and chondroitin sulfatases in Alzheimer's Disease were reviewed. Measurements of SAA2, iNOS, lipid peroxidation, chondroitin sulfate proteoglycan 4, and other parameters were performed in cortex and hippocampus from ARSB-null mice and controls by QRT-PCR, ELISA, and other standard assays.
Results: SAA2 mRNA expression and protein, CSPG4 mRNA, chondroitin 4-sulfate and i-NOS were increased significantly in ARSB-null mice. Measures of lipid peroxidation and redox state were significantly modified.
Discussion: Findings indicate that decline in ARSB leads to changes in expression of parameters associated with AD in the hippocampus and cortex of the ARSB-deficient mouse.
Conclusions: Further investigation of the impact of decline in ARSB on the development of AD may provide a new approach to prevent and treat AD.

DOI: https://doi.org/10.1101/2023.04.03.535377
J Biol Chem. 2023 Apr 13. pii: S0021-9258(23)01741-6. [Epub ahead of print] 104713

TFCP2 is a transcriptional regulator of heparan sulfate assembly and melanoma cell growth.

Amrita Basu, Rachel N Champagne, Neil G Patel, Elijah D Nicholson, Ryan J Weiss.

  Heparan sulfate (HS) is a long, linear polysaccharide that is ubiquitously expressed in all animal cells and plays a key role in many cellular processes, including cell signaling and development. Dysregulation of HS assembly has been implicated in pathophysiological conditions, such as tumorigenesis and rare genetic disorders. HS biosynthesis occurs in a non-template driven manner in the endoplasmic reticulum and Golgi through the activity of a large group of biosynthetic enzymes. While much is known about its biosynthesis, little is understood about the regulation of HS assembly across diverse tissue types and disease states. To address this gap in knowledge, we recently performed genome-wide CRISPR/Cas9 screens to identify novel regulatory factors of HS biosynthesis. From these screens, we identified the alpha globin transcription factor, TFCP2, as a top hit. To investigate the role of TFCP2 in HS assembly, we targeted TFCP2 expression in human melanoma cells using the CRISPR/Cas9 system. TFCP2 knockout cells exhibited decreased fibroblast growth factor binding to cell surface HS, alterations in HS composition, and slowed cell growth compared to wildtype cells. Additionally, RNA sequencing revealed that TFCP2 regulates expression of multiple enzymes involved in HS assembly, including the secreted endosulfatase, SULF1. Pharmacological targeting of TFCP2 activity similarly reduced growth factor binding and increased SULF1 expression, and knockdown of SULF1 expression in TFCP2 mutant cells restored melanoma cell growth. Overall, these studies identify TFCP2 as a novel transcriptional regulator of HS and highlight HS-protein interactions as a possible target to slow melanoma growth.

Keywords:  TFCP2; gene regulation; glycosaminoglycans; heparan sulfate; melanoma; sulfatase 1; transcription factor

DOI:  https://doi.org/10.1016/j.jbc.2023.104713
Cell Rep Phys Sci. 2023 Apr 19. 4(4): 101346

SARS-CoV-2 evolved variants optimize binding to cellular glycocalyx.

Sang Hoon Kim, Fiona L Kearns, Mia A Rosenfeld, Lane Votapka, Lorenzo Casalino, Micah Papanikolas, Rommie E Amaro, Ronit Freeman.

  Viral variants of concern continue to arise for SARS-CoV-2, potentially impacting both methods for detection and mechanisms of action. Here, we investigate the effect of an evolving spike positive charge in SARS-CoV-2 variants and subsequent interactions with heparan sulfate and the angiotensin converting enzyme 2 (ACE2) in the glycocalyx. We show that the positively charged Omicron variant evolved enhanced binding rates to the negatively charged glycocalyx. Moreover, we discover that while the Omicron spike-ACE2 affinity is comparable to that of the Delta variant, the Omicron spike interactions with heparan sulfate are significantly enhanced, giving rise to a ternary complex of spike-heparan sulfate-ACE2 with a large proportion of double-bound and triple-bound ACE2. Our findings suggest that SARS-CoV-2 variants evolve to be more dependent on heparan sulfate in viral attachment and infection. This discovery enables us to engineer a second-generation lateral-flow test strip that harnesses both heparin and ACE2 to reliably detect all variants of concern, including Omicron.

Keywords:  ACE2; Brownian dynamics; COVID-19; SARS-CoV-2; biomimetic; biosensor; electrostatic potential; heparan sulfate; heparin; lateral-flow assay; spike

DOI:  https://doi.org/10.1016/j.xcrp.2023.101346
Eur Thyroid J. 2023 Apr 01. pii: ETJ-23-0023. [Epub ahead of print]

Novel (sulfated) thyroid hormone transporters in the solute carrier 22 family.

Zhongli Chen, Robin P Peeters, Wesley Flach, Linda J de Rooij, Sena Yildiz, Alexander Teumer, Matthias Nauck, Rosalie B T M Sterenborg, Joost H W Rutten, Marco Medici, W Edward Visser, Marcel E Meima.

OBJECTIVE: Thyroid hormone (TH) transport represents a critical first step in governing intracellular TH regulation. It is still unknown whether the full repertoire of TH transporters has been identified. Members of the solute carrier (SLC) 22 family have substrates in common with the known TH transporters of the organic anion transporting peptide (OATP) family. Therefore we screened the SLC22 family for TH transporters.
METHODS: Uptake of 1 nM of iodothyronines or sulfated iodothyronines in COS1 cells expressing SLC22 proteins was performed.
RESULTS: We first tested 25 mouse (m) SLC22 proteins for TH uptake and found that the majority of the organic anion transporter (OAT) clade were capable of 3,3',5-triiodothyronine (T3) and/or thyroxine (T4) transport. Based on phylogenetic tree analysis of the mouse and human (h) SLC22 family, we selected 8 hSLC22s that grouped with the newly identified mouse TH transporters. Of these, 4 tested positive for uptake of one or more substrates, particularly hSLC22A11 showed robust (3-fold over control) uptake of T4. Uptake of sulfated iodothyronines was strongly (up to 17-fold) induced by some SLC22s, most notably SLC22A8, hSLC22A9, mSLC22A27 and mSLC22A29. Finally, the zebrafish orthologues of SLC22A6/8 drOatx and drSlc22a6l also transported almost all (sulfated) iodothyronines tested. The OAT-inhibitors lesinurad and probenecid inhibited most SLC22 proteins.
CONCLUSIONS: Our results demonstrated that members of the OAT clade of the SLC22 family constitute a novel, evolutionary conserved group of transporters for (sulfated) iodothyronines. Future studies should reveal the relevance of these transporters in TH homeostasis and physiology.

DOI: https://doi.org/10.1530/ETJ-23-0023
Front Endocrinol (Lausanne). 2023 ;14 1081056

Acute changes in serum and skeletal muscle steroids in resistance-trained men.

Felipe C Vechin, Jakob L Vingren, Guilherme D Telles, Miguel S Conceicao, Cleiton A Libardi, Manoel E Lixandrao, Felipe Damas, Telma F Cunha, Patricia C Brum, Luiz A Riani, Carlos Ugrinowitsch.

   Introduction: Resistance exercise can significantly increase serum steroid concentrations after an exercise bout. Steroid hormones are involved in the regulation of several important bodily functions (e.g., muscle growth) through both systemic delivery and local production. Thus, we aimed to determine whether resistance exercise-induced increases in serum steroid hormone concentrations are accompanied by enhanced skeletal muscle steroid concentrations, or whether muscle contractions per se induced by resistance exercise can increase intramuscular steroid concentrations.
Methods: A counterbalanced, within-subject, crossover design was applied. Six resistance-trained men (26 ± 5 years; 79 ± 8 kg; 179 ± 10 cm) performed a single-arm lateral raise exercise (10 sets of 8 to 12 RM - 3 min rest between sets) targeting the deltoid muscle followed by either squat exercise (10 sets of 8 to 12 RM - 1 min rest) to induce a hormonal response (high hormone [HH] condition) or rest (low hormone [LH] condition). Blood samples were obtained pre-exercise and 15 min and 30 min post-exercise; muscle specimens were harvested pre-exercise and 45 min post-exercise. Immunoassays were used to measure serum and muscle steroids (total and free testosterone, dehydroepiandrosterone sulfate, dihydrotestosterone, and cortisol; free testosterone measured only in serum and dehydroepiandrosterone only in muscle) at these time points.
Results: In the serum, only cortisol significantly increased after the HH protocol. There were no significant changes in muscle steroid concentrations after the protocols.
Discussion: Our study provides evidence that serum steroid concentration increases (cortisol only) seem not to be aligned with muscle steroid concentrations. The lack of change in muscle steroid after protocols suggests that resistance-trained individuals were desensitized to the exercise stimuli. It is also possible that the single postexercise timepoint investigated in this study might be too early or too late to observe changes. Thus, additional timepoints should be examined to determine if RE can indeed change muscle steroid concentrations either by skeletal muscle uptake of these hormones or the intramuscular steroidogenesis process.

Keywords:  intracrine; intracrinology; measurement error; steroidogenesis; strength exercise

DOI:  https://doi.org/10.3389/fendo.2023.1081056
Exp Eye Res. 2023 Apr 15. pii: S0014-4835(23)00097-0. [Epub ahead of print] 109476

The role of KS GAGs in the microstructure of CXL-treated corneal stroma; a transmission electron microscopy study.

H Hatami-Marbini, M E Emu.

  The mechanical and physical properties of the cornea originate from the microstructure and composition of its extracellular matrix. It is known that collagen fibrils, with a relatively uniform diameter, are organized in a pseudo-hexagonal array. It has been suggested that proteoglycans and the interaction of their glycosaminoglycan (GAG) side chains with themselves and collagen fibrils are important for collagen fibril organization inside the cornea. There are several diseases such as keratoconus in which the regular collagen fibrillar packing becomes distorted causing corneal optical and mechanical properties to be compromised. The primary purpose of the present work was to investigate the role of GAGs on the microstructure of corneal extracellular matrix before and after corneal collagen crosslinking (CXL). For this purpose, keratan sulphates (KS) were removed from corneal samples using the keratanase enzyme and the CXL procedure was used to crosslink the specimens. The transmission electron microscopy was then used to characterize the diameter of collagen fibrils and their interfibrillar spacing. It was found that KS GAG depletion increased the collagen interfibrillar spacing while the CXL treatment significantly decreased the interfibrillar spacing. The enzyme and CXL treatment had an insignificant effect on the diameter of collagen fibrils. The underlying mechanisms responsible for these observations were discussed in terms of the assumption that GAG chains form duplexes that behave as tiny ropes holding collagen fibrils in place.

Keywords:  Collagen fibril diameter; Corneal extracellular matrix; Glycosaminoglycans; TEM; Ultrastructure

DOI:  https://doi.org/10.1016/j.exer.2023.109476
Int J Biol Macromol. 2023 Apr 18. pii: S0141-8130(23)01396-X. [Epub ahead of print] 124502

Analyzing the effect of heparin on in vitro capacitation and spermatozoal RNA population in goats.

Bijayalaxmi Sahoo, Balaram Mishra, Rakesh Bhaskar, Y N V Vikas, Anushri Umesh, Praveen Kumar Guttula, Mukesh Kumar Gupta.

  Heparin is a glycosaminoglycan polymer that is commonly used as an anticoagulant. Heparin also induces in vitro capacitation in spermatozoa, although its molecular mechanism is elusive. This study investigated the effect of heparin on in vitro capacitation and spermatozoal RNA (spRNA) population in goats. Goat spermatozoa were treated with 20 μM heparin for 0-6 h and evaluated for motility, capacitation, acrosome reaction, and spRNA population by RNA sequencing (RNA-seq). It was observed that heparin enhanced sperm motility up to 6 h of incubation (p < 0.05). Heparin also induced capacitation and acrosome reaction within 4 h. RNA-seq identified 1254 differentially expressed genes (DEGs) between heparin-treated and control spermatozoa. Most DEGs (1251 nos.) were upregulated and included 1090 protein-coding genes. A few genes (PRND, ITPR1, LLCFC1, and CHRM2) showed >5-fold increased expression in heparin-treated spermatozoa compared to the control. The upregulated genes were found to be involved in cAMP-PKA, PI3-Akt, calcium, MAPK signaling, and oxidative stress pathways. DCFDA staining confirmed the increased oxidative stress in heparin-treated spermatozoa compared to the control (p < 0.05). In conclusion, the results of the present study suggest that heparin enhances sperm motility and induces capacitation by upregulation of the spRNA population and oxidative stress pathway.

Keywords:  Capacitation; Goat; Heparin; RNA-seq; Spermatozoal RNA; Transcriptome

DOI:  https://doi.org/10.1016/j.ijbiomac.2023.124502