| 1. |
- Faustini, Francesca, et al.
(författare)
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First exposure to rituximab is associated to high rate of anti-drug antibodies in systemic lupus erythematosus but not in ANCA-associated vasculitis
- 2021
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Ingår i: Arthritis Research & Therapy. - : BMC. - 1478-6362. ; 23:1
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Tidskriftsartikel (refereegranskat)abstract
- Background Anti-drug antibodies (ADAs) can impact on the efficacy and safety of biologicals, today used to treat several chronic inflammatory conditions. Specific patient groups may be more prone to develop ADAs. Rituximab is routinely used for ANCA-associated vasculitis (AAV) and as off-label therapy for systemic lupus erythematosus (SLE), but data on occurrence and predisposing factors to ADAs in these diseases is limited. Objectives To elucidate the rate of occurrence, and risk factors for ADAs against rituximab in SLE and AAV. Methods ADAs were detected using a bridging electrochemiluminescent (ECL) immunoassay in sera from rituximab-naive (AAV; n = 41 and SLE; n = 62) and rituximab-treated (AAV; n = 22 and SLE; n = 66) patients. Clinical data was retrieved from medical records. Disease activity was estimated by the SLE Disease Activity Index-2000 (SLEDAI-2 K) and the Birmingham Vasculitis Activity Score (BVAS). Results After first rituximab cycle, no AAV patients were ADA-positive compared to 37.8% of the SLE patients. Samples were obtained at a median (IQR) time of 5.5 (3.7-7.0) months (AAV), and 6.0 (5.0-7.0) months (SLE). ADA-positive SLE individuals were younger (34.0 (25.9-40.8) vs 44.3 (32.7-56.3) years, p = 0.002) and with more active disease (SLEDAI-2 K 14.0 (10.0-18.5) vs. 8.0 (6.0-14), p = 0.0017) and shorter disease duration (4.14 (1.18-10.08) vs 9.19 (5.71-16.93), p = 0.0097) compared to ADA-negative SLE. ADAs primarily occurred in nephritis patients, were associated with anti-dsDNA positivity but were not influenced by concomitant use of corticosteroids, cyclophosphamide or previous treatments. Despite overall reduction of SLEDAI-2 K (12.0 (7.0-16) to 4.0 (2.0-6.7), p < 0.0001), ADA-positive individuals still had higher SLEDAI-2 K (6.0 (4.0-9.0) vs 4.0 (2.0-6.0), p = 0.004) and their B cell count at 6 months follow-up was higher (CD19 + % 4.0 (0.5-10.0) vs 0.5 (0.4-1.0), p = 0.002). At retreatment, two ADA-positive SLE patients developed serum sickness (16.7%), and three had infusion reactions (25%) in contrast with one (5.2%) serum sickness in the ADA-negative group. Conclusions In contrast to AAV, ADAs were highly prevalent among rituximab-treated SLE patients already after the first course of treatment and were found to effect on both clinical and immunological responses. The high frequency in SLE may warrant implementations of ADA screening before retreatment and survey of immediate and late-onset infusion reactions.
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| 2. |
- Faustini, Francesca
(författare)
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Studies on biological treatment and biomarkers in systemic lupus erythematosus : longterm effects, risk factors and predictors of response
- 2021
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Systemic Lupus Erythematosus (SLE) is an autoimmune disease in which several immune mechanisms are involved in a complex interplay. B-cells are considered to be a major player in the pathophysiology of the disease, and the advent of B-cell depletion therapy (BCDT), has brought much promise. Yet, after twenty years of use of the depleting agent Rituximab (RTX), BCDT still has to find its precise location and role in SLE therapy. Among SLE clinical manifestations, lupus nephritis (LN) carries a significant burden in terms of morbidity, and requires optimised approaches. Identifying clinical and biological biomarkers, may contribute to improving the clinical management of the disease and its major organ involvement. The aims of this thesis were the following: 1) to contribute to a definition of RTX’s role in SLE treatment, by studying its deep immunological effects, immunogenicity and side effects, aiming at identifying possible biomarkers of efficacy and safety which might be implemented in daily practice; 2) to identify possible non-invasive biomarkers of specific organ involvement, such as LN, which may also in the near future be implemented in daily practice for identifying LN, and to monitor disease activity and response to treatment. In Paper I, deep immunological effects of RTX on recently defined B- and T- cell subsets were explored, through a multicolour flow cytometry approach. In particular, we investigated whether RTX affects the age-associated B-cells (ABCs) which are plasma cells precursors; and the T-follicular and T-peripheral helper T-cells (TFH, TPH). We showed that transient reduction of the frequencies of the B-cell phenotype age-associated B-cells (ABCs) is induced by the treatment during the early phases of B-cell depletion. This corresponded to a reduction within the DN2 compartment, which contains the ABCs cells. By contrast, no early significant changes of the TFH and TPH followed the administration of RTX. A reduction of a subset of CD4+ with high expression of the marker PD-1 was shown at later follow-up. Examining the behaviour of these cell subsets in patients who developed antibodies against the drug, we showed that immunogenicity was associated to lower frequencies of double negative (DN) B-cells and to a wider expansion of plasma blasts at early follow up. In Paper II, we investigated the occurrence of anti-drug antibodies (ADAs) towards RTX, in a cohort of 66 SLE patients after their first course of treatment, in comparison with 22 first-ever treated ANCA-associated vasculitis (AAV) patients. After the first course of RTX almost 38% of SLE patients developed ADA, while in AAV no ADA were shown. The SLE patients developing ADA were younger, had a longer disease duration, and a more active disease at the time of treatment initiation. They were mostly treated for LN and had a serologically active profile. Despite an overall reduction of disease activity upon RTX treatment, the presence of ADA in SLE was associated with higher counts of B-cells in the peripheral blood at the 6 month follow-up, and with higher residual disease activity. At retreatment, ADA-positive SLE patients experienced more frequent infusion reactions, both of immediate and late-onset occurrence. In Paper III, we investigated the occurrence of late onset neutropenia (LON) in 107 patients with SLE treated with RTX. We found a rate of occurrence as high as 30%. In most cases the laboratory finding was discovered in routine controls and did not cause complications. However, around 40% of the patients experienced clinical consequences with occurrence of neutropenic fever and infections, some requiring intensive care. The clinical characteristic associated with the risk of LON was high disease activity before treatment. At a biological level, the appearance of LON was associated with the increase of BAFF which follows the disappearance of B-cells in the peripheral blood. In Paper IV, we explored the use of the soluble protein Galectin-3 Binding Protein (Gal- 3BP), as urinary biomarker in LN, comparing the urinary concentration of this protein in 86 patients with active LN, 63 patients with active SLE without LN, 73 inactive SLE patients, and 48 matched population based controls. Samples were tested separately for the concentrations of other biomarkers: Galectin-3 (Gal-3), neutrophil gelatinase associated lipocalin (NGAL), osteopontin (OPN) and kidney injury molecule-1 (KIM-1). Each of the biomarkers was assessed as absolute concentration and as concentration normalized for the urine-creatinine (adjusted concentration). We found that the levels of both non-adjusted and adjusted u-Gal-3BP were significantly higher in the urine of patients with active LN as compared to all the other groups. Higher levels of adjusted u-Gal-3BP were found in proliferative and membranous forms of LN as compared to mesangial forms. We also found a moderate correlation between the adjusted u-Gal-3BP and the histological activity index as evaluated in kidney biopsies. This correlation was stronger when we considered patients not receiving immunosuppressive treatments at the time of kidney biopsy. Current treatment with oral corticosteroids was associated with lower urinary levels of Gal-3BP in active LN patients. In patients with proliferative LN this association was found also regarding ongoing treatment with antimalarials. In a subset of ten patients with active LN, significant reduction of Gal-3BP levels was observed at repeated analysis after treatment. In Paper V, we explored the presence in the urine of LN patients (n=13) of extracellular vesicles (EVs) carrying an array of molecules of interest, all with a putative role in inflammation. We found that all the tested EVs were detectable in the urine, although at lower concentrations as compared to blood. We preliminary adopted a cut-off of EVs concentration of 50x10^6/L and evaluated the concentration of EVs carrying the cargo molecules of interest above the above mentioned cut-off. We considered the expression of EVs with respect to the histological activity. No correlation was found, but the concentration of EVs carrying the split complement molecule C5a, was found to be significantly higher in patients classified as active at renal biopsy as compared to those classified as inactive. We then examined the expression of urinary EVs with respect to having a predominant proliferative histologic pattern with respect to non-proliferative pattern. In proliferative LN, urinary EVs expressing C3a, C4, C4d, C5a, Mitochondrial antigens, Lactadherin, NGAL and TWEAK were found in significantly higher concentrations as compared to patients with non-proliferative forms. In conclusion, the thesis here presented adds some elements of knowledge on the immunological effects and consequences of the use of RTX in SLE, and on aspects of safety associated with immunogenicity. It also adds knowledge on the magnitude of LON as a safety aspect of RTX use as a therapy for SLE. Finally, it contributes to the definition of potential urinary biomarkers which might be implemented in clinical practice in the future.
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| 3. |
- Faustini, Francesca, et al.
(författare)
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Urine Galectin-3 binding protein reflects nephritis activity in systemic lupus erythematosus
- 2023
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Ingår i: Lupus. - : Sage Publications. - 0961-2033 .- 1477-0962. ; 32:2, s. 252-262
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Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND: Lupus nephritis (LN) is a major and severe organ involvement in systemic lupus erythematosus (SLE), whose diagnosis and treatment necessitate to perform kidney biopsy, which is an invasive procedure. Non-invasive urine biomarkers are an active area of investigation to support LN diagnosis and management.OBJECTIVE: To investigate the role of urinary galectin-3 binding protein (u-Gal-3BP) as a candidate biomarker of renal disease in biopsy proven LN.PATIENTS AND METHODS: Levels of u-Gal-3BP were investigated in a cross-sectional fashion by ELISA in 270 subjects: 86 LN patients, 63 active SLE patients with no kidney involvement, 73 SLE patients with inactive disease and 48 age and sex-matched population-based controls (PBC). Moreover, urine samples were analysed separately by ELISA for additional markers of kidney pathology: neutrophil gelatinase-associated lipocalin (NGAL), osteopontin (OPN), kidney injury molecule-1 (KIM-1) and galectin-3 (Gal-3). The concentrations of all studied molecules were normalized to urine creatinine levels. In 10 patients, post-treatment levels of the biomarkers were measured.RESULTS: Normalized u-Gal-3BP levels were higher in LN patients compared to the other groups (p < .0001). Comparing different LN classes, u-Gal-3BP levels were higher among patients with proliferative (class III/IV) and membranous (class V) as compared to mesangial (class II) forms (p = .04). In proliferative forms, u-Gal-3BP levels correlated with the activity index in renal biopsies (r = 0.42, p = .004). Moreover, in a subset of 10 patients with repeated kidney biopsy and urine sampling before and after induction treatment, a significant decrease of u-Gal-3BP was observed (p = .03). Among the other markers, KIM-1 was also able to discriminate LN from the other groups, while NGAL, OPN and Gal-3 could not in this cohort.CONCLUSION: Given its ability to discriminate LN patients from active non-renal and inactive SLE patients, the observed correlation with the activity index in renal biopsies, and its levels declining following treatment, u-Gal-3BP shows promise as a non-invasive urinary biomarker to help detecting and to monitor renal involvement in SLE patients and should be validated in larger cohorts.
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| 4. |
- Faustini, Gaia, et al.
(författare)
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Synapsin III deficiency hampers α-synuclein aggregation, striatal synaptic damage and nigral cell loss in an AAV-based mouse model of Parkinson’s disease
- 2018
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Ingår i: Acta Neuropathologica. - : Springer Science and Business Media LLC. - 0001-6322 .- 1432-0533. ; 136:4, s. 621-639
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Tidskriftsartikel (refereegranskat)abstract
- Parkinson’s disease (PD), the most common neurodegenerative movement disorder, is characterized by the progressive loss of nigral dopamine neurons. The deposition of fibrillary aggregated α-synuclein in Lewy bodies (LB), that is considered to play a causative role in the disease, constitutes another key neuropathological hallmark of PD. We have recently described that synapsin III (Syn III), a synaptic phosphoprotein that regulates dopamine release in cooperation with α-synuclein, is present in the α-synuclein insoluble fibrils composing the LB of patients affected by PD. Moreover, we observed that silencing of Syn III gene could prevent α-synuclein fibrillary aggregation in vitro. This evidence suggests that Syn III might be crucially involved in α-synuclein pathological deposition. To test this hypothesis, we studied whether mice knock-out (ko) for Syn III might be protected from α-synuclein aggregation and nigrostriatal neuron degeneration resulting from the unilateral injection of adeno-associated viral vectors (AAV)-mediating human wild-type (wt) α-synuclein overexpression (AAV-hαsyn). We found that Syn III ko mice injected with AAV-hαsyn did not develop fibrillary insoluble α-synuclein aggregates, showed reduced amount of α-synuclein oligomers detected by in situ proximity ligation assay (PLA) and lower levels of Ser129-phosphorylated α-synuclein. Moreover, the nigrostriatal neurons of Syn III ko mice were protected from both synaptic damage and degeneration triggered by the AAV-hαsyn injection. Our observations indicate that Syn III constitutes a crucial mediator of α-synuclein aggregation and toxicity and identify Syn III as a novel therapeutic target for PD.
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| 5. |
- Faustini, Gaia, et al.
(författare)
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Synapsin III gene silencing redeems alpha-synuclein transgenic mice from Parkinsons disease-like phenotype
- 2022
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Ingår i: Molecular Therapy. - : Cell Press. - 1525-0016 .- 1525-0024. ; 30:4, s. 1465-1483
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Tidskriftsartikel (refereegranskat)abstract
- Fibrillary aggregated alpha-synuclein (alpha-syn) deposition in Lewy bodies (LB) characterizes Parkinsons disease (PD) and is believed to trigger dopaminergic synaptic failure and a retrograde terminal-to-cell body neuronal degeneration. We described that the neuronal phosphoprotein synapsin III (Syn III) cooperates with alpha-syn to regulate dopamine (DA) release and can be found in the insoluble alpha-syn fibrils composing LB. Moreover, we showed that a-syn aggregates deposition, and the associated onset of synaptic deficits and neuronal degeneration occurring following adeno-associated viral vectors-mediated overexpression of human alpha-syn in the nigrostriatal system are hindered in Syn III knock out mice. This supports that Syn III facilitates alpha-syn aggregation. Here, in an interventional experimental design, we found that by inducing the gene silencing of Syn III in human alpha-syn transgenic mice at PD-like stage with advanced alpha-syn aggregation and overt striatal synaptic failure, we could lower alpha-syn aggregates and striatal fibers loss. In parallel, we observed recovery from synaptic vesicles clumping, DA release failure, and motor functions impairment. This supports that Syn III consolidates alpha-syn aggregates, while its downregulation enables their reduction and redeems the PD-like phenotype. Strategies targeting Syn III could thus constitute a therapeutic option for PD.
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| 6. |
- Häyry, Aliisa, et al.
(författare)
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Interleukin (IL) 16 : a candidate urinary biomarker for proliferative lupus nephritis
- 2022
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Ingår i: Lupus Science and Medicine. - : BMJ Publishing Group Ltd. - 2053-8790. ; 9:1
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Tidskriftsartikel (refereegranskat)abstract
- OBJECTIVE: Lupus nephritis (LN) is a severe manifestation of systemic lupus erythematosus (SLE). The pathogenesis is incompletely understood and diagnostic biomarkers are scarce. We investigated interleukin (IL) 16 as a potential biomarker for LN in a well-characterised cohort of patients with SLE.METHODS: We measured urinary (u-) and plasma (p-) levels of IL-16 in predefined patient groups using ELISA: LN (n=84), active non-renal SLE (n=63), inactive non-renal SLE (n=73) and matched population controls (n=48). The LN group included patients with recent biopsy-confirmed proliferative (PLN, n=47), mesangioproliferative (MES, n=11) and membranous (MLN, n=26) LN. Renal expression of IL-16 was investigated by immunohistochemistry. Associations between IL-16 measurements and clinical parameters and the diagnostic value for LN were explored.RESULTS: p-IL-16 was detected in all investigated cases and high p-IL-16 levels were observed in patients with active SLE. u-IL-16 was detected (dt-u-IL-16) in 47.6% of patients with LN, while only up to 17.8% had dt-u-IL-16 in other groups. In the LN group, 68% of patients with PLN had dt-u-IL-16, while the proportions in the MLN and MES groups were lower (11.5% and 45.5%, respectively). The highest u-IL-16 levels were detected in the PLN group. In the regression model, u-IL-16 levels differentiated PLN from other LN patient subgroups (area under the curve 0.775-0.896, p<0.0001). dt-u-IL-16 had superior specificity but slightly lower sensitivity than elevated anti-double-stranded DNA and low complement C3 or C4 in diagnosing PLN. A high proportion of LN kidney infiltrating cells expressed IL-16.CONCLUSIONS: We demonstrate that detectable u-IL-16 can differentiate patients with PLN from those with less severe LN subtypes and active non-renal SLE. Our findings suggest that u-IL-16 could be used as a screening tool at suspicion of severe LN. Furthermore, the high IL-16 levels in plasma, urine and kidney tissue imply that IL-16 could be explored as a therapeutic target in SLE.
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| 7. |
- Ostergaard, Mikkel, et al.
(författare)
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Certolizumab pegol, abatacept, tocilizumab or active conventional treatment in early rheumatoid arthritis : 48-week clinical and radiographic results of the investigator-initiated randomised controlled NORD-STAR trial
- 2023
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Ingår i: Annals of the Rheumatic Diseases. - : BMJ PUBLISHING GROUP. - 0003-4967 .- 1468-2060. ; 82:10, s. 1286-1295
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Tidskriftsartikel (refereegranskat)abstract
- Background The optimal first-line treatment in early rheumatoid arthritis (RA) is debated. We compared clinical and radiographic outcomes of active conventional therapy with each of three biological treatments with different modes of action. Methods Investigator-initiated, randomised, blinded-assessor study. Patients with treatment-naive early RA with moderate-severe disease activity were randomised 1:1:1:1 to methotrexate combined with (1) active conventional therapy: oral prednisolone (tapered quickly, discontinued at week 36) or sulfasalazine, hydroxychloroquine and intra-articular glucocorticoid injections in swollen joints; (2) certolizumab pegol; (3) abatacept or (4) tocilizumab. Coprimary endpoints were week 48 Clinical Disease Activity Index (CDAI) remission (CDAI <= 2.8) and change in radiographic van der Heijde-modified Sharp Score, estimated using logistic regression and analysis of covariance, adjusted for sex, anticitrullinated protein antibody status and country. Bonferroni's and Dunnet's procedures adjusted for multiple testing (significance level: 0.025). Results Eight hundred and twelve patients were randomised. Adjusted CDAI remission rates at week 48 were: 59.3% (abatacept), 52.3% (certolizumab), 51.9% (tocilizumab) and 39.2% (active conventional therapy). Compared with active conventional therapy, CDAI remission rates were significantly higher for abatacept (adjusted difference +20.1%, p<0.001) and certolizumab (+13.1%, p=0.021), but not for tocilizumab (+12.7%, p=0.030). Key secondary clinical outcomes were consistently better in biological groups. Radiographic progression was low, without group differences. Conclusions Compared with active conventional therapy, clinical remission rates were superior for abatacept and certolizumab pegol, but not for tocilizumab. Radiographic progression was low and similar between treatments.
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