Protocolo de valoración de patología articular en paleopatología: estudio de la rodilla en la necrópolis hispanomusulmana de San Nicolás (Murcia, s. XI-XIII)

Trabajo presentado para obtener el Diploma de Estudios Avanzados (DEA)

El sistema mononuclear fagocítico en la progresión de la artritis reumatoide y de la artrosis

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Bioquímica. Fecha de lectura: 28 marzo, 2014La artritis reumatoide (AR) y la artrosis (OA) son dos enfermedades reumáticas que comparten mecanismos de lesión tisular, aun siendo de etiopatogenia completamente diferente. Mientras que la sinovitis persistente es el distintivo principal de la AR, hallazgos experimentales y clínicos sugieren que la inflamación sinovial puede estar asociada con la progresión de la OA. Las células del sistema mononuclear fagocítico (SMF) desempeñan múltiples y cruciales funciones en la sinovitis, a pesar de lo cual se desconocen otros aspectos relacionados con su diferenciación y su contribución al daño tisular. Entre ellos, el fenotipo de las células multinucleadas gigantes, ocasionalmente descritas en la sinovitis de la AR y la OA. En este trabajo describimos la elevada frecuencia con la que están presentes estas células, la proximidad tisular de los diversos detritus que probablemente estimulan su diferenciación, así como la asociación con la intensidad de la respuesta inflamatoria. Las células del SMF interaccionan con las diferentes estirpes celulares presentes en la sinovial inflamada, tanto las residentes como las provenientes de la sangre. En este sentido, nos preguntamos si otras células extra-sinoviales pero con participación en la patogenia de ambas artropatías crónicas pudieran tener relación con el SMF. Hemos demostrado que el condrocito puede contribuir, mediante la producción de RANKL por el cartílago y, posterior incremento de la osteoclastogénesis subcondral, a la osteopenia yuxta-articular de la AR. Los factores de riesgo cardiovascular tradicionales, junto con la inflamación sistémica, empeoran la progresión de la arteriosclerosis en la AR. Hemos observado que uno de los componentes del síndrome metabólico, la hipercolesterolemia, también empeora la progresión de la sinovitis reumatoide y la destrucción periarticular erosiva por un incremento de la osteoclastogénesis. En conjunto, estos resultados resaltan la importancia del SMF en la artritis crónica siendo una diana potencial para el tratamiento de la sinovitis en AR y en OA.Rheumatoid arthritis (RA) and osteoarthritis (OA) are two rheumatic diseases sharing tissue lesions mechanisms, despite of their completely different etiopathogenesis. While persistent synovitis is the main hallmark of RA, experimental and clinical findings suggest that synovial inflammation may be associated with OA progression. Mononuclear phagocyte system (MPS) cells take on multiple and crucial functions within the inflamed synovium, however other aspects of its differentiation and contribution to tissue damage remain poorly understood. Among them, multinucleated giant cell (MGC) phenotype which has occasionally been described in rheumatoid and osteoarthritis synovitis. In this study, we first aimed to describe the high frequency with which these cells are present, the close proximity to specific tissue debris probably stimulating its differentiation and its association with the intensity of the inflammatory response. The cells of the MPS interplay with a large variety of synovial cells, both resident and resulting from blood. We wonder whether other extra-synovial cells, also participating in the pathogenesis of both chronic arthropathies, could be associated with the MPS. We have demonstrated how articular chondrocytes may contribute to yuxta-articular bone loss in RA, by producing RANKL in cartilage and subsequently increasing subchondral osteoclastogenesis. The increase of traditional cardiovascular risk factors, along with systemic inflammation, impairs atherosclerosis in RA. We have observed how one of the components of the metabolic syndrome, hyperlipidemia, impairs rheumatoid synovitis progression and periarticular erosive destruction by increasing osteoclastogenesis. Overall results point out the relevance of the MPS in chronic arthritis suggesting that these cells are a potential target for reducing synovitis in RA and OA

Characterization of multinucleated giant cells in synovium and subchondral bone in knee osteoarthritis and rheumatoid arthritis

Background: Multinucleated giant cells have been noticed in diverse arthritic conditions since their first description in rheumatoid synovium. However, their role in the pathogenesis of osteoarthritis (OA) or rheumatoid arthritis (RA) still remains broadly unknown. We aimed to study the presence and characteristics of multinucleated giant cells (MGC) both in synovium and in subchondral bone tissues of patients with OA or RA. Methods: Knee synovial and subchondral bone samples were from age-matched patients undergoing total joint replacement for OA or RA, or non-arthritic post mortem (PM) controls. OA synovium was stratified by histological inflammation grade using index tissue sections. Synovitis was assessed by Krenn score. Histological studies employed specific antibodies against macrophage markers or cathepsin K, or TRAP enzymatic assay. Results: Inflamed OA and RA synovia displayed more multinucleated giant cells than did non-inflamed OA and PM synovia. There was a significant association between MGC numbers and synovitis severity. A TRAP negative/cathepsin K negative Langhans-like subtype was predominant in OA, whereas both Langhans-like and TRAP-positive/ cathepsin K negative foreign-body-like subtypes were most commonly detected in RA. Plasma-like and foam-like subtypes also were observed in OA and RA synovia, and the latter was found surrounding adipocytes. TRAP positive/ cathepsin K positive osteoclasts were only identified adjacent to subchondral bone surfaces. TRAP positive osteoclasts were significantly increased in subchondral bone in OA and RA compared to PM controls. Conclusions: Multinucleated giant cells are associated with synovitis severity, and subchondral osteoclast numbers are increased in OA, as well as in RA. Further research targeting multinucleated giant cells is warranted to elucidate their contributions to the symptoms and joint damage associated with arthritis

Mediators and Patterns of Muscle Loss in Chronic Systemic Inflammation

Besides its primary function in locomotion, skeletal muscle (SKM), which represents up to half of human's weight, also plays a fundamental homeostatic role. Through the secretion of soluble peptides, or myokines, SKM interacts with major organs involved in metabolic processes. In turn, metabolic cues from these organs are received by muscle cells, which adapt their response accordingly. This is done through an intricate intracellular signaling network characterized by the cross-talking between anabolic and catabolic pathways. A fine regulation of the network is required to protect the organism from an excessive energy expenditure. Systemic inflammation evokes a catabolic reaction in SKM known as sarcopenia. In turn this response comprises several mechanisms, which vary depending on the nature of the insult and its magnitude. In this regard, aging, chronic inflammatory systemic diseases, osteoarthritis and idiopathic inflammatory myopathies can lead to muscle loss. Interestingly, sarcopenia may persist despite remission of chronic inflammation, an issue which warrants further research. The Janus kinase/signal transducer and activator of transcription (JAK/STAT) system stands as a major participant in muscle loss during systemic inflammation, while it is also a well-recognized orchestrator of muscle cell turnover. Herein we summarize current knowledge about models of sarcopenia, their triggers and major mediators and their effect on both protein and cell growth yields. Also, the dual action of the JAK/STAT pathway in muscle mass changes is discussed. We highlight the need to unravel the precise contribution of this system to sarcopenia in order to design targeted therapeutic strategies

Additional file 1: of Characterization of multinucleated giant cells in synovium and subchondral bone in knee osteoarthritis and rheumatoid arthritis

Morphological MGC foam subtype and CD68 positive cells surrounding fat cells in OA and in RA. A and B. MGCs displaying a foam-like subtype were identified near to and surrounding fat cells in inflamed synovia from patients with either OA (A) or RA (B). Haematoxylin and eosin staining. Scale bar = 20 μm. Open arrows indicate foam-like MGC and A = adipocyte. C and D. Multiple mononuclear CD68 positive cells were found in a crown-like structure encircling adipocyte cells in both OA (C) and RA (D). Immunohistochemistry for CD68, using eosin contrast. Scale bar = 100 μm. (TIFF 3238 kb

Compensatory anabolic signaling in the sarcopenia of experimental chronic arthritis

Inflammatory activity in rheumatoid arthritis may alter the regulation of muscle mass leading to a secondary sarcopenia, commonly termed rheumatoid cachexia (RC). We characterized alterations to muscle structure and various pro-inflammatory, catabolic and regenerative markers in an animal model of RC. Antigen induced arthritis (AiA) was performed in 20 male adult rabbits. AiA animals exhibited significantly less weight gain, a markedly elevated serum C-reactive protein (CRP), lighter muscles with shorter cross-sectional diameter and increased myonuclei when compared to controls. Atrogin-1 and MuRF-1 were up-regulated alongside an increase in IL-1β, active NF-κB and a higher ratio of phosphorylated to inactive p38 MAPK. CCL-2 and TNF levels were reduced and IL-6 was unchanged between groups. We observed decreased pSTAT3, unchanged pSTAT1 and Myf5, but increased Pax7, MyoD and myogenin. AiA rabbits had a reduction in myostatin from gastrocnemii and synovium with a congruent decrease in serum myostatin compared to controls. Chronic arthritis induced an RC-like secondary sarcopenia with increased muscle protein breakdown. Elevated IL-1β may trigger proteolysis via elevated NF-κB and p38 MAPK signaling with a compensatory anabolic response suggested by myonuclear expansion, increased Pax7, MyoD and myogenin, reduced pSTAT3 as well as reduced serum, synovial and muscular myostatinThis work was partially supported by research grants from the Instituto de Salud Carlos III (PI13/00570, PI15/00340, PI16/00065 and RETICEF RD12/0043/0008), co-funded by Fondo Europeo de Desarrollo Regional (FEDER) and ASPIRE IIR grant from Pfizer

Comprehensive Approach to Genomic and Immune Profiling: Insights of a Real-World Experience in Gynecological Tumors

Gynecological cancer accounts for an elevated incidence worldwide requiring responsiveness regarding its care. The comprehensive genomic approach agrees with the classification of certain tumor types. We evaluated 49 patients with gynecological tumors undergoing high-throughput sequencing to explore whether identifying alterations in cancer-associated genes could characterize concrete histological subtypes. We performed immune examination and analyzed subsequent clinical impact. We found 220 genomic aberrations mostly distributed as single nucleotide variants (SNV, 77%). Only 3% were classified as variants of strong clinical significance in BRCA1 and BRCA2 of ovarian high-grade serous (HGSC) and uterine endometrioid carcinoma. TP53 and BRCA1 occurred in 72% and 28% of HGSC. Cervical squamous cell carcinoma was entirely HPV-associated and mutations occurred in PIK3CA (60%), as well as in uterine serous carcinoma (80%). Alterations were seen in PTEN (71%) and PIK3CA (60%) of uterine endometrioid carcinoma. Elevated programmed death-ligand 1 (PD-L1) was associated with high TILs. Either PD-L1 augmented in deficient mis-matched repair (MMR) proteins or POLE mutated cases when compared to a proficient MMR state. An 18% received genotype-guided therapy and a 4% immunotherapy. The description of tumor subtypes is plausible through high-throughput sequencing by recognizing clinically relevant alterations. Additional concomitant assessment of immune biomarkers identifies candidates for immunotherapy

Multicenter evaluation of the Idylla NRAS-BRAF mutation test in metastatic colorectal cancer

Treatment of colorectal cancer (CRC) with monoclonal antibodies against epidermal growth factor receptor requires the assessment of the mutational status of exons 2, 3, and 4 of the NRAS and KRAS oncogenes. Moreover, the mutational status of exon 15 of the BRAF oncogene is a marker of poor prognosis in CRC. The Idylla NRAS-BRAF Mutation Test is a reliable, simple (<2 minutes hands-on time), and quick (<2 hours turnaround time) sample-to-result solution, enabling the detection of clinically relevant mutations in NRAS (18 mutations) and BRAF (5 mutations). A multicenter study was conducted in 14 centers using the Idylla NRAS-BRAF Mutation Test to assess the NRAS and BRAF mutational status of 418 formalin-fixed, paraffin-embedded tissue samples from CRC patients. Results were compared with those obtained earlier by routine reference methods, including next-generation sequencing, pyrosequencing, mass spectrometry-based assays, PCR-based assays, and Sanger sequencing. In case of discordance, additional tests were performed by digital droplet PCR. Overall, after testing confirmation and excluding invalids/errors by design, concordances between the Idylla NRAS-BRAF Mutation Test and the reference test results were found in almost perfect agreement. In conclusion, the Idylla NRAS-BRAF Mutation Test enables the routine detection of all NRAS and BRAF mutations deemed clinically relevant according to the latest clinical guidelines, without necessitating molecular expertise or infrastructure