Studies to Assess the Utility of Serum Neurofilament Light Chain as a Biomarker in Chemotherapy-Induced Peripheral Neuropathy

Chemotherapy-induced peripheral neuropathy (CIPN) is one of the most common and disabling dose-limiting toxicities of chemotherapy. We report here the results of two separate non-interventional studies (49 patients), which evaluated blood neurofilament light chain (NfL) as a biomarker of CIPN in breast cancer patients treated with paclitaxel. All patients underwent a standard treatment protocol that was established independently of the present studies. NfL was measured in serum using an ultrasensitive single-molecule array and compared with the self-administered European Organization for Research and Treatment of Cancer Quality of Life Questionnaire-CIPN twenty-item scale (CIPN20) and Total Neuropathy Score clinical version (TNSc), a clinician-reported measure of neuropathy progression. The TNSc increased with cumulative dose compared with baseline, and the NfL concentrations were also strongly associated with the cumulative dose of chemotherapy. The analysis showed a correlation between TNSc and NfL. Both TNSc and NfL showed weak to moderate associations with CIPN20 subscores, with a better association for the CIPN20 sensory compared with motor and autonomic subscores. Data from the two studies provide evidence that serum NfL has the potential to be used as a biomarker to monitor and mitigate CIPN. However, studies with additional patients planned in the ongoing clinical trial will determine the universal application of NfL as a biomarker in CIPN

Oxidative Stress in Non-Alcoholic Fatty Liver Disease

Non-alcoholic fatty liver disease (NAFLD) is a challenging disease caused by multiple factors, which may partly explain why it still remains an orphan of adequate therapies. This review highlights the interaction between oxidative stress (OS) and disturbed lipid metabolism. Several reactive oxygen species generators, including those produced in the gastrointestinal tract, contribute to the lipotoxic hepatic (and extrahepatic) damage by fatty acids and a great variety of their biologically active metabolites in a “multiple parallel-hit model”. This leads to inflammation and fibrogenesis and contributes to NAFLD progression. The alterations of the oxidant/antioxidant balance affect also metabolism-related organelles, leading to lipid peroxidation, mitochondrial dysfunction, and endoplasmic reticulum stress. This OS-induced damage is at least partially counteracted by the physiological antioxidant response. Therefore, modulation of this defense system emerges as an interesting target to prevent NAFLD development and progression. For instance, probiotics, prebiotics, diet, and fecal microbiota transplantation represent new therapeutic approaches targeting the gut microbiota dysbiosis. The OS and its counter-regulation are under the influence of individual genetic and epigenetic factors as well. In the near future, precision medicine taking into consideration genetic or environmental epigenetic risk factors, coupled with new OS biomarkers, will likely assist in noninvasive diagnosis and monitoring of NAFLD progression and in further personalizing treatments

Exploring serum and CSF Calcitonin Gene Related Peptide levels: A promising biomarker in multiple sclerosis?

Introduction: Calcitonin Gene Related Peptide (CGRP) is a neuropeptide ubiquitous in the peripheral and central nervous system, mostly known for the role in vasodilation and pain signal transmission during migraine attacks. Recent studies have been unraveling its immunomodulatory properties, including its possible role in multiple sclerosis (MS) pathophysiology, however there is no conclusive evidence on whether it plays a pro or anti-inflammatory role. Objectives/Aims: To evaluate soluble CGRP levels at MS diagnosis, in cerebrospinal fluid (CSF) and serum, and evaluate associations with progression and short-term disease severity. Methods: We enrolled for a retrospective cohort study 59 patients (39 females, mean age at diagnosis 38.79 years ± standard deviation or SD 9.89) with Radiological Isolated Syndrome (RIS), Clinical Isolated Syndrome (CIS) and Relapsing-Remitting (RR) MS. During the diagnostic work-up were collected clinic-demographic data, serum and CSF. Patients were followed with clinical visits in which clinical data were collected.*** CGRP levels were determined through an ELISA commercial kit (MyBioSource Inc, MBS267126, San Diego, CA, USA). None had a history of migraine attack at diagnosis. Statistical analyses were conducted with STATA software to determine Mann–Whitney, Kruskal-Wallis test and Spearman’s rank correlation coefficient significance. Results: CGRP levels were significantly higher in MS patients if compared to healthy controls published by Papiri et Al. (PMID: 37013432) and Han et Al. (PMID: 35204700). Mean values resulted 73.10 pg/ml in serum (±9.42 vs 29.50 ± 8.91, p<0.05 t-test) and 64.01 in CSF (± 10.39 vs 52.05 ± 5.70, p<0.05 t-test). CGRP levels did not relate to clinical variables at diagnosis: age, gender, Expanded Disability Status Scale (EDSS), number of T2, gadolinium enhancing and spinal cord lesions. However, there was a positive correlation between serum CGRP and the Multiple Sclerosis Severity Score (MSSS) at the last follow up (r2 = 0.27, p<0.05 Spearman’s rank correlation). Conclusion: We observed an increased CGRP level in the CSF and serum of MS patients at diagnosis. Our findings suggest its potential use as a biomarker to identify cases with poor prognosis, indicating a pro-inflammatory effect of this neuropeptide

Longitudinal Assessment of Transorbital Sonography, Visual Acuity, and Biomarkers for Inflammation and Axonal Injury in Optic Neuritis

Background and Objective. To investigate the relationship between optic nerve sheath diameter, optic nerve diameter, visual acuity and osteopontin, and neurofilament heavy chain in patients with acute optic neuritis. Patients and Methods. Sonographic and visual acuity assessment and biomarker measurements were executed in 23 patients with unilateral optic neuritis and in 19 sex- and age-matched healthy controls. Results. ONSD was thicker on the affected side at symptom onset (median 6.3 mm; interquartile range 6.0–6.5) than after 12 months (5.3 mm; 4.9–5.6; p<0.001) or than in controls (5.2 mm; 4.8–5.5; p<0.001). OND was significantly increased in the affected side (3.4 mm; 2.9–3.8) compared to healthy controls (2.7 mm; 2.5–2.9; p<0.001) and was thicker at baseline than after 12 months (2.8 mm; 2.7–3.0; p<0.01). Visual acuity improved significantly after 12 months (1.00; 0.90–1.00) compared to onset of symptoms (0.80; 0.40–1.00; p<0.001). OPN levels were significantly higher in patients at presentation (median 6.44 ng/ml; 2.05–10.06) compared to healthy controls (3.21 ng/ml, 1.34–4.34; p<0.03). Concentrations of NfH were significantly higher in patients than in controls. Conclusion. ONSD and OND are increased in the affected eye. OPN and NfH are elevated in patients, confirming the presence of any underlying inflammation and axonal injury

Probiotics May Have Beneficial Effects in Parkinson's Disease: In vitro Evidence

Background: Parkinson's disease (PD) is characterized by loss of dopaminergic neurons and intraneuronal accumulation of alpha-synuclein, both in the basal ganglia and in peripheral sites, such as the gut. Peripheral immune activation and reactive oxygen species (ROS) production are important pathogenetic features of PD. In this context, the present study focused on the assessment of in vitro effects of probiotic bacterial strains in PBMCs isolated from PD patients vs. healthy controls.Methods: 40 PD patients and 40 matched controls have been enrolled. Peripheral blood mononuclear cells (PBMCs) were isolated and co-cultured with a selection of probiotics microorganisms belonging to the lactobacillus and bifidobacterium genus. In vitro release of the major pro- (Tumor Necrosis Factor-alpha and Interleukin-17A and 6) and anti-inflammatory (Interleukin 4 and 10) cytokines by PBMCs, as well as the production of ROS was investigated. Furthermore, we assessed the ability of probiotics to influence membrane integrity, antagonize the growth of potential pathogen bacteria, such as Escherichia coli and Klebsiella pneumoniae and encode tyrosine decarboxylase genes (tdc).Results: All probiotic strains were able to inhibit inflammatory cytokines and ROS production in both patients and controls. The most striking results were obtained in PD subjects with L. salivarius LS01 and L. acidophilus which significantly reduced pro-inflammatory and increased the anti-inflammatory cytokines (p &lt; 0.05). Furthermore, most strains determined restoration of membrane integrity and inhibition of E. coli and K. pneumoniae. Finally, we also showed that all the strains do not carry tdc gene, which is known to decrease levodopa bioavailability in PD patients under treatment.Conclusions: Probiotics exert promising in vitro results in decreasing pro-inflammatory cytokines, oxidative stress and potentially pathogenic bacterial overgrowth. In vivo longitudinal data are mandatory to support the use of bacteriotherapy in PD

Gas6/TAM system: potential prognostic biomarker for Multiple Sclerosis

Introduction: The protein growth arrest specific 6 (Gas6) and its tyrosine kinase receptors Tyro-3, Axl, Mer (TAMs) are ubiquitous proteins involved in regulation of inflammation and apoptotic body clearance. Gas6 and TAMs have been associated with neuronal remyelination and stimulation of oligodendrocyte survival. However, few data are available on their role in multiple sclerosis (MS). Objectives/Aims: Objectives/Aims: In this study we evaluated if soluble levels of these molecules, determined at MS diagnosis in cerebrospinal fluid (CSF) and serum, correlated with progression with short-term disease severity. Methods: Methods: We conducted a retrospective cohort study enrolling 64 patients with different forms of MS, the Radiological Isolated Syndrome (RIS), the Clinical Isolated Syndrome (CIS) and Relapsing-Remitting (RR). At diagnosis, we collected serum, CSF, and clinical-radiological data: lesion load, spinal cord, and gadolinium-enhancing (Gad+) lesions, and expanded disability status score (EDSS). During the last clinical follow-up EDSS, MS severity score (MSSS) and Age-Related MS severity (ARMSS) were assessed. Gas6 and TAMs were determined by ELISA kit (R&D Systems), while neurofilaments (NFLs) levels, for neuronal damage assessment, by SimplePlexTM fluorescence-based immunoassay. Statistical analyses were conducted with STATA software to determine Mann–Whitney, Kruskal-Wallis test and Spearman’s rank correlation coefficient significance. Results: Results: At diagnosis, RIS and CIS showed higher values of sMer and sTyro-3, compared to RRMS (p = 0.007 and p = 0.018). Serum sAxl was higher in patients untreated or first-line disease modifying treatments (DMTs) versus patients with high-efficacy DMTs (p = 0.04). Moreover, serum Axl was associated with EDSS ≤ 3 at diagnosis (p = 0.037) and EDSS progression in patients with EDSS ≤ 3 (p = 0.017). Similarly, high levels of Gas6 in CSF were associated with EDSS ≤ 3 at diagnosis (p = 0.04), and high levels of Gas6 in serum to a lower MSSS (r2 = -0.32 and p = 0.01). Results significances were confirmed by multivariate analyses. In our cohort, serum and CSF NFLs levels were confirmed as markers of disability in EDSS (p = 0.005 and p = 0.002) and MSSS (r2 = 0.27 and p =0.03; r2 = 0.39 and p = 0.001). Conclusion: Conclusion: Taken together, our results suggest that Gas6 and its receptors, particularly Axl, might have a neuroprotective role and prognostic potential in MS. Disclosures: Disclosures: Nothing to disclos

Extracellular proteasome-osteopontin circuit regulates cell migration with implications in multiple sclerosis

Osteopontin is a pleiotropic cytokine that is involved in several diseases including multiple sclerosis. Secreted osteopontin is cleaved by few known proteases, modulating its pro-inflammatory activities. Here we show by in vitro experiments that secreted osteopontin can be processed by extracellular proteasomes, thereby producing fragments with novel chemotactic activity. Furthermore, osteopontin reduces the release of proteasomes in the extracellular space. The latter phenomenon seems to occur in vivo in multiple sclerosis, where it reflects the remission/relapse alternation. The extracellular proteasome-mediated inflammatory pathway may represent a general mechanism to control inflammation in inflammatory diseases

Experimental Models to Study Immune Dysfunction in the Pathogenesis of Parkinson’s Disease

Parkinson’s disease (PD) is a chronic, age-related, progressive multisystem disease associated with neuroinflammation and immune dysfunction. This review discusses the methodological approaches used to study the changes in central and peripheral immunity in PD, the advantages and limitations of the techniques, and their applicability to humans. Although a single animal model cannot replicate all pathological features of the human disease, neuroinflammation is present in most animal models of PD and plays a critical role in understanding the involvement of the immune system (IS) in the pathogenesis of PD. The IS and its interactions with different cell types in the central nervous system (CNS) play an important role in the pathogenesis of PD. Even though culture models do not fully reflect the complexity of disease progression, they are limited in their ability to mimic long-term effects and need validation through in vivo studies. They are an indispensable tool for understanding the interplay between the IS and the pathogenesis of this disease. Understanding the immune-mediated mechanisms may lead to potential therapeutic targets for the treatment of PD. We believe that the development of methodological guidelines for experiments with animal models and PD patients is crucial to ensure the validity and consistency of the results

Longitudinal assessment of transorbital sonography, visual acuity, and biomarkers for inflammation and axonal injury in optic neuritis

BACKGROUND AND OBJECTIVE: To investigate the relationship between optic nerve sheath diameter, optic nerve diameter, visual acuity and osteopontin, and neurofilament heavy chain in patients with acute optic neuritis. PATIENTS AND METHODS: Sonographic and visual acuity assessment and biomarker measurements were executed in 23 patients with unilateral optic neuritis and in 19 sex- and age-matched healthy controls. RESULTS: ONSD was thicker on the affected side at symptom onset (median 6.3\u2009mm; interquartile range 6.0-6.5) than after 12 months (5.3\u2009mm; 4.9-5.6; p < 0.001) or than in controls (5.2\u2009mm; 4.8-5.5; p < 0.001). OND was significantly increased in the affected side (3.4\u2009mm; 2.9-3.8) compared to healthy controls (2.7\u2009mm; 2.5-2.9; p < 0.001) and was thicker at baseline than after 12 months (2.8\u2009mm; 2.7-3.0; p < 0.01). Visual acuity improved significantly after 12 months (1.00; 0.90-1.00) compared to onset of symptoms (0.80; 0.40-1.00; p < 0.001). OPN levels were significantly higher in patients at presentation (median 6.44\u2009ng/ml; 2.05-10.06) compared to healthy controls (3.21\u2009ng/ml, 1.34-4.34; p < 0.03). Concentrations of NfH were significantly higher in patients than in controls. CONCLUSION: ONSD and OND are increased in the affected eye. OPN and NfH are elevated in patients, confirming the presence of any underlying inflammation and axonal injury