Juvenile neuropsychiatric systemic lupus erythematosus: identification of novel central neuroinflammation biomarkers

International audienceIntroduction Juvenile systemic lupus erythematosus (j-SLE) is a rare chronic autoimmune disease affecting multiple organs. Ranging from minor features, such as headache or mild cognitive impairment, to serious and life-threatening presentations, j-neuropsychiatric SLE (j-NPSLE) is a therapeutic challenge. Thus, the diagnosis of NPSLE remains difficult, especially in pediatrics, with no specific biomarker of the disease yet validated. Objectives To identify central nervous system (CNS) disease biomarkers of j-NPSLE. Methods A 5-year retrospective tertiary reference monocentric j-SLE study. A combination of standardized diagnostic criteria and multidisciplinary pediatric clinical expertise was combined to attribute NP involvement in the context of j-SLE. Neopterin and interferon-alpha (IFN-α) protein levels in cerebrospinal fluid (CSF) were assessed, together with routine biological and radiological investigations. Results Among 51 patients with j-SLE included, 39% presented with j-NPSLE. J-NPSLE was diagnosed at onset of j-SLE in 65% of patients. No specific routine biological or radiological marker of j-NPSLE was identified. However, CSF neopterin levels were significantly higher in active j-NPSLE with CNS involvement than in j-SLE alone ( p = 0.0008). Neopterin and IFN-α protein levels in CSF were significantly higher at diagnosis of j-NPSLE with CNS involvement than after resolution of NP features (respectively p = 0.0015 and p = 0.0010) upon immunosuppressive treatment in all patients tested ( n = 10). Both biomarkers correlated strongly with each other ( R s = 0.832, p < 0.0001, n = 23 paired samples). Conclusion CSF IFN-α and neopterin constitute promising biomarkers useful in the diagnosis and monitoring of activity in j-NPSLE

Proteomic profiling of a mouse model for ovarian granulosa cell tumor identifies VCP as a highly sensitive serum tumor marker in several human cancers.

The initial aim of this study was to identify novel serum diagnostic markers for the human ovarian granulosa cell tumor (GCT), a tumor that represents up to 5% of all ovarian cancers. To circumvent the paucity of human tissues available for analyses, we used the Ctnnb1(tm1Mmt/+);Pten(tm1Hwu/tmiHwu);Amhr2(tm3(cre)Bhr/+) transgenic mouse model, which features the constitutive activation of CTNNB1 signaling combined with the loss of Pten in granulosa cells and develops GCTs that mimic aggressive forms of the human disease. Proteomic profiling by mass spectrometry showed that vinculin, enolase 1, several heat shock proteins, and valosin containing protein (VCP) were more abundantly secreted by cultured mouse GCT cells compared to primary cultured GC. Among these proteins, only VCP was present in significantly increased levels in the preoperative serum of GCT cancer patients compared to normal subjects. To determine the specificity of VCP, serum levels were also measured in ovarian carcinoma, non-Hodgkin's lymphoma and breast, colon, pancreatic, lung, and prostate cancer patients. Increased serum VCP levels were observed in the majority of cancer cases, with the exception of patients with lung or prostate cancer. Moreover, serum VCP levels were increased in some GCT, ovarian carcinoma, breast cancer, and colon cancer patients who did not otherwise display increased levels of widely used serum tumor markers for their cancer type (e.g. inhibin A, inhibin B, CA125, CEA, or CA15.3). These results demonstrate the potential use of VCP as highly sensitive serum marker for GCT as well as several other human cancers

Serum levels of GCT markers in healthy women or in women with GCT.

<p>Measurements that exceeded the normal reference range are indicated in bold. For inhibin A and B (INHA and INHB), values beneath the detection thresholds by ELISA were defined as normal. For VCP, the reference value was set as the mean of healthy control band intensity in immunoblot analyses+2SD (253 arbitrary units). Note that serum inhibin usually becomes undetectable after menopause in healthy women. Interpretation of premenopausal inhibin values can be difficult due to their secretion both by growing ovarian follicles and by GCTs. NE: not evaluated.</p

Mass spectrometry analysis of the secretome of mouse GCT from CPA transgenic mice.

<p>Protein identification (International Protein Index identifier) and protein description are given along with the overall score and the number of peptides identified by mass spectrometry as described in Materials & Methods. Proteins in bold were studied further.</p

Tumor clinical features and VCP serum level in tested cancer patients.

<p>For VCP, the reference value was set as the mean of healthy control band intensity in immunoblot analyses+2SD (253 arbitrary units). Positive VCP values are indicated in bold. X: unknown grade/differentiation, CL: clear cell, EM: endometroid, NOS: not otherwise specified, IDC: invasive ductal carcinoma, LC: large cell, NHDG: non-Hodgkin's lymphoma, NA: not available.</p

Assessment of serum levels of VCP compared to serum tumor markers currently used for ovarian carcinoma, colon cancer, and breast cancer.

<p>Sera from patients were tested for VCP levels by immunoblot analyses and the dotted line shows VCP cutoff values established in healthy donors. In addition, sera were tested by ELISA for the presence (+) or absence (−) of increased levels of CA125 in ovarian carcinoma, CEA in colon cancer, or CEA and CA15.3 in breast cancer. The normal ranges of CA125, CEA, and CA15.3 are below 35 U/ml, 7 µg/l, and 29 kU/l, respectively.</p

Differential protein expression in mouse granulosa cell and GCT cell culture media.

<p>Samples were separated by 10% SDS-PAGE followed by silver staining. Arrows indicate examples of proteins expressed selectively in the GCT cell culture media, along with approximate molecular weights. A total of 3 gels with different acrylamide contents were studied and 14 bands (with proteins from 12 to 116 KDa) were subjected to mass spectrometry analysis. Only one of the 3 gels is shown on the figure.</p

Respiratory complications in pediatric epidermal necrolysis: A retrospective study of 22 cases

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