Role of the HSP70 Co-Chaperone SIL1 in Health and Disease

Cell surface and secreted proteins provide essential functions for multicellular life. They enter the endoplasmic reticulum (ER) lumen co-translationally, where they mature and fold into their complex three-dimensional structures. The ER is populated with a host of molecular chaperones, associated co-factors, and enzymes that assist and stabilize folded states. Together, they ensure that nascent proteins mature properly or, if this process fails, target them for degradation. BiP, the ER HSP70 chaperone, interacts with unfolded client proteins in a nucleotide-dependent manner, which is tightly regulated by eight DnaJ-type proteins and two nucleotide exchange factors (NEFs), SIL1 and GRP170. Loss of SIL1′s function is the leading cause of Marinesco-Sjögren syndrome (MSS), an autosomal recessive, multisystem disorder. The development of animal models has provided insights into SIL1′s functions and MSS-associated pathologies. This review provides an in-depth update on the current understanding of the molecular mechanisms underlying SIL1′s NEF activity and its role in maintaining ER homeostasis and normal physiology. A precise understanding of the underlying molecular mechanisms associated with the loss of SIL1 may allow for the development of new pharmacological approaches to treat MSS.     

Rose bengal staining as an assessment of ocular surface damage and recovery in dry eye disease—A review

PurposeTo review the use of ophthalmic rose bengal to assess the ocular surface, especially considering the grading scales used.MethodsA literature search was undertaken to identify reports where either rose bengal or lissamine green had been used as a ‘vital’ stain, with a special interest in identifying studies that provided data before and after treatment for dry eye and also considered the mechanism of action of these two chemicals.ResultsBetween 1985 and 2006, numerous clinical studies used a consistent grading scheme (that attributed to Van Bijsterveld) to assess the outcome of artificial tear treatments on dry eye patients. With such consistency, including the adoption of a treatment period of 1 month, comparisons can be made to indicate the efficacy of rose bengal staining to assess reduction in ocular surface desiccation. However, in the following years, several alternative grading schemes have been used for both rose bengal and lissamine green and assessment periods have been variable so making inter-study comparisons considerably more difficult to undertake. An attribute of rose bengal appears to be its ability to stain the nuclei of cells, but whether this also occurs for lissamine green is unclear.ConclusionsOphthalmic rose bengal has been successfully adopted for use to assess the ocular surface over many years as a vital stain. More research is needed to assess whether lissamine green ocular surface staining can simply be substituted for rose bengal in evaluation of dry eye treatments.     

Toll-Like Receptor 7 Is Required for Lacrimal Gland Autoimmunity and Type 1 Diabetes Development in Male Nonobese Diabetic Mice

Sjögren syndrome (SS) is an immunologically complex, chronic autoimmune disease targeting lacrimal and salivary glands. Nonobese diabetic (NOD) mice spontaneously develop inflammation of lacrimal and salivary glands with histopathological features similar to SS in humans including focal lymphocytic infiltrates in the affected glands. The innate immune signals driving lymphocytic infiltration of these glands are not well-defined. Here we evaluate the role of Toll-like receptor (TLR) 7 in the development of SS-like manifestations in NOD mice. We created a Tlr7 knockout NOD mouse strain and performed histological and gene expression studies to characterize the effects of TLR7 on autoimmunity development. TLR7 was required for male-specific lacrimal gland inflammation but not for female-specific salivary gland inflammation. Moreover, TLR7 was required for type 1 diabetes development in male but not female NOD mice. RNA sequencing demonstrated that TLR7 was associated with a type I interferon (IFN) response and a type I IFN-independent B cell response in the lacrimal glands. Together these studies identify a previously unappreciated pathogenic role for TLR7 in lacrimal gland autoimmunity and T1D development in male NOD mice adding to the growing body of evidence supporting sex differences in mechanisms of autoimmune disease in NOD mice.     

Treatment of Sjögren’s Syndrome with Mesenchymal Stem Cells: A Systematic Review

Mesenchymal stem cells (MSCs) are ubiquitous in the human body. Mesenchymal stem cells were initially isolated from bone marrow and later from other organs such as fatty tissues, umbilical cords, and gingiva. Their secretory capacities give them interesting immunomodulatory properties in cell therapy. Some studies have explored the use of MSCs to treat Sjögren’s syndrome (SS), a chronic inflammatory autoimmune disease that mainly affects exocrine glands, including salivary and lacrimal glands, although current treatments are only palliative. This systematic review summarizes the current data about the application of MSCs in SS. Reports show improvements in salivary secretions and a decrease in lymphocytic infiltration in salivary glands in patients and mice with SS after intravenous or infra-peritoneal injections of MSCs. MSC injections led to a decrease in inflammatory cytokines and an increase in anti-inflammatory cytokines. However, the intrinsic mechanism of action of these MSCs currently remains unknown.     

Salivary and Lacrimal Gland Alterations of the Epidermal Fatty Acid-Binding Protein (E-FABP) in Non-Obese Diabetic Mice

The purpose of this study was to investigate the changes in E-FABP in the salivary and lacrimal glands of the Sjögren syndrome (SS) model non-obese diabetic mice (NOD). Cotton thread and ocular vital staining tests were performed on 10-week NOD male mice (n = 24) and age- and sex-matched wild-type (WT) mice (n = 25). Tear and saliva samples were collected at sacrifice for E-FABP ELISA assays. Salivary and lacrimal gland specimens underwent immunohistochemistry stainings for E-FABP. Real-time RT-PCR was also performed for the quantification of mRNA expression levels in the salivary and lacrimal glands. Corneal vital staining scores in the NOD mice were significantly higher compared with those for the wild-type mice (p = 0.0001). The mean tear E-FABP level showed a significantly lower concentration in the NOD mice (p = 0.001). The mean saliva E-FABP level also showed a significantly lower concentration in the NOD mice (p = 0.04). Immunohistochemistry revealed intense E-FABP staining in the LG acinar epithelium and less intense staining in the acinar epitheliae of the SGs in the NOD mice compared to the WT mice. Real-time RT-PCR for the mRNA expression of E-FABP showed a significantly decreased expression in the SG and a significant increase in the LG of the NOD mice compared to the WT mice. In conclusion, the E-FABP showed marked alterations in the tear film, saliva, lacrimal, and salivary glands of the NOD mouse, which may help explain the ocular surface changes in relation to the dry eye disease in this SS model mouse and keratoconjunctivitis sicca in SS patients.     

Decreased BAFF Receptor Expression and Unaltered B Cell Receptor Signaling in Circulating B Cells from Primary Sjögren’s Syndrome Patients at Diagnosis

Animal models of autoimmunity and human genetic association studies indicate that the dysregulation of B-cell receptor (BCR) signaling is an important driver of autoimmunity. We previously showed that in circulating B cells from primary Sjögren’s syndrome (pSS) patients with high systemic disease activity, protein expression of the BCR signaling molecule Bruton’s tyrosine kinase (BTK) was increased and correlated with T-cell infiltration in the target organ. We hypothesized that these alterations could be driven by increased B-cell activating factor (BAFF) levels in pSS. Here, we investigated whether altered BCR signaling was already present at diagnosis and distinguished pSS from non-SS sicca patients. Using (phospho-)flow cytometry, we quantified the phosphorylation of BCR signaling molecules, and investigated BTK and BAFF receptor (BAFFR) expression in circulating B cell subsets in an inception cohort of non-SS sicca and pSS patients, as well as healthy controls (HCs). We found that both BTK protein levels and BCR signaling activity were comparable among groups. Interestingly, BAFFR expression was significantly downregulated in pSS, but not in non-SS sicca patients, compared with HCs, and correlated with pSS-associated alterations in B cell subsets. These data indicate reduced BAFFR expression as a possible sign of early B cell involvement and a diagnostic marker for pSS.     

Tear Proteomics Approach to Distinguishing Primary from Secondary Sjögren’s Syndrome for Dry Eye Patients with Long-Term Instillation of Eyedrops

The diagnosis and monitoring of Sjögren syndrome (SS) is often difficult, requiring a multidisciplinary approach with invasive procedures. Our aim is to elucidate the tear protein alterations of dry eye disease (DED) with primary SS (pSS) and secondary SS (sSS) with the long-term instillation of eyedrops. We collected clinical demographics and tear fluid (TF) samples from DED patients with no autoimmune diseases (non-SS-DED), pSS-DED, and sSS-DED patients, followed by TF screening with tandem mass tagging-labeling gel-free proteomics assay. Bioinformatic analysis via Ingenuity Pathway Analysis was used to identify functional pathways and interacting networks. Validation of candidate proteins with enzyme-linked immunosorbent assay on the tear samples was done. The top functional pathways of the two comparisons (sSS-DED vs. pSS-DED and sSS-DED vs. non-SS-DED) were both associated with inflammation and stress-related signaling. After constructing an interaction network model with the selected candidate proteins, five proteins were identified. A Disintegrin and Metalloproteinase domain-containing protein 10 (ADAM10) was found to be an important candidate biomarker in all groups, followed by epidermal growth factor (EGF) in TF. This study revealed novel DED markers, ADAM10 and EGF, in differentiating between primary and secondary SS patients from tears by in-depth proteomic analysis.     

Association between symptoms of xerostomia and dry eye in older people

PurposeTo explore the association between xerostomia and dry eye symptoms in the older adult population.MethodsSecondary analysis of data from 627 participants (346 male, 291 female), with a mean ± SD age of 75 ± 7 years, recruited through the population-based South Australian Dental Longitudinal Study. Participants completed the validated Summated Xerostomia Inventory–Dutch Version questionnaire (SXI-D), and rated their overall frequency of dry eye symptoms at baseline.ResultsSymptomatic dry eye was reported by 31% of participants, and xerostomia by 21%. A positive correlation was observed between dry eye symptoms and SXI-D scores (Spearman’s rho = +0.379, p < 0.001). Overall, participants with symptomatic dry eye had higher SXI-D scores than those without (10.6 ± 3.6 vs. 8.1 ± 2.8, p < 0.001), and were more likely to report xerostomia, with an odds ratio (95% CI) of 2.25 (1.52–3.35; p < 0.001).ConclusionsSymptoms of xerostomia and dry eye were relatively common in the cohort of older adult participants. The potentially debilitating implications of – and positive association between – the two types of sicca symptoms support the routine evaluation of xerostomia symptoms as part of the assessment of dry eye patients, and vice versa.     

Multicenter review of impression-based scleral devices

ObjectiveTo describe outcomes using impression based-scleral devices for the management of anterior segment disease.MethodsRetrospective chart review identified all patients who were fitted with impression-based scleral devices between January 1, 2013 and June 30, 2019 at three specialty contact lens practices. Patient demographic data, indication for device use, visual and physiological outcomes, as well as details of the fitting process and survival of device use were determined.ResultsForty-four patients (70 eyes) were included in the study. Primary indications for device use included corneal irregularity (28 patients, 44 eyes) and ocular surface disease (16 patients, 26 eyes). Fifty-four percent of patients had more than one ocular surface condition, and 39 % of patients had undergone at least one anterior segment surgical procedure. Twenty-nine patients had unsuccessfully attempted to wear standard scleral lenses prior to being fit with impression-based devices. Visual acuity improved significantly with impression-based devices compared to habitual correction (p < 0.001). Completion of the fitting process (including visit to acquire the impression and post-fitting assessments) required an average of 4 [1.5] visits.Ideal haptic alignment was achieved with 74 % and complete limbal clearance was achieved in 83 % of fits. Device use was discontinued due to complications in two eyes.ConclusionPatients with complex eye disease who are unable to successfully wear standard scleral lenses successfully may achieve visual and therapeutic success with impression-based devices.