Anti-thymic stromal lymphopoietin monoclonal antibody in patients with chronic rhinosinusitis with nasal polyps (DUBHE): Rationale and design of a multicenter, randomized, double-blind, placebo-controlled study.

Background: Chronic rhinosinusitis with nasal polyps (CRSwNP) has a complex pathogenesis and is difficult to treat, which brings a huge economic burden to society. Despite all the progress in the treatment of CRSwNP, some patients with CRSwNP still experience recurrence. Therefore, there is an urgent need to develop novel drugs and treatments for CRSwNP. Thymic stromal lymphopoietin (TSLP) is produced by epithelial cells and mediates type 2 and nontype 2 inflammation through various downstream cellular immune and inflammatory pathways. Anti-TSLP treatment with tezepelumab has been proven to be effective in treating patients with uncontrolled asthma, regardless of their peripheral blood eosinophil levels being low or high. However, there is no relevant research on the usage of anti-TSLP monoclonal antibodies for the treatment of uncontrolled CRSwNP. Objective: This is the first phase Ib/IIa study for subjects with uncontrolled CRSwNP, aiming to evaluate the safety, tolerability, pharmacokinetics, pharmacodynamics, immunogenicity, and preliminary efficacy of multiple ascending doses (MAD) of anti-TSLP monoclonal antibody. Methods: The DUBHE is a multicenter, randomized, double-blind, placebo-controlled, phase Ib/IIa clinical study. The study will be composed of 3 periods: a screening/run-in period of 4 weeks, a treatment period of 52 weeks (16 weeks of double-blind treatment period +36 weeks of open-label treatment period), and a safety follow-up period of 12 weeks. No more than 113 subjects with uncontrolled CRSwNP will be divided into 4 groups to receive different doses of CM326 or placebo treatments (55 mg every two weeks [Q2W] group, 110 mg Q2W group, 220 mg Q2W group, and 220 mg every four weeks [Q4W] group). Enrolled patients will be stratified by tissue eosinophil count (TEC). Results: The safety of the monoclonal antibody that targets TSLP in uncontrolled CRSwNP and its preliminary efficacy at 16 weeks of treatment. Conclusion: In this study, for the first time, the safety and preliminary efficacy of MAD of CM326 will be verified. The efficacy of CM326 in patients with eosinophilic CRSwNP (TEC ≥55/high power field [HPF]), as well as noneosinophilic CRSwNP (TEC <55/HPF) will be testified. Trial registration: NCT05324137.

Microvessel barrier dysfunction in sinonasal inverted papilloma-associated squamous cell carcinoma and its manifestation in dynamic contrast-enhanced MRI.

BACKGROUND: To date, an effective means to preoperatively predict the malignant transformation of sinonasal inverted papilloma (SIP) remains lacking due to similarities in clinical appearance. This study aimed to retrospectively evaluate dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) parameters and microvessel structure in tumors with histologically confirmed SIP and inverted papilloma-associated squamous cell carcinoma (IP-SCC), as well as correlate DCE-MRI findings with angiogenesis biomarkers. METHODS: Absolute quantitative DCE-MRI parameters (Ktrans, Kep, Ve) based on the Tofts model and model-free semi-quantitative indices (Tpeak, WR, MaxSlope) of SIP (n = 22) and IP-SCC (n = 20) were investigated. Regions of interest (ROIs) were oriented according to the tumor subsites in the surgical records. Micro-vessel density (MVD) counts and tight junction protein (claudin-5) expression were evaluated in tumor specimens obtained during surgery. Differences in the above data were compared between the two groups. Correlations between DCE-MRI parameters and angiogenic biomarkers were analyzed. RESULTS: Compared with SIP specimens, IP-SCC specimens were characterized by a significantly higher MVD and a leakier microvessel barrier. The values of Tpeak and Ve were significantly higher for SIP than those for IP-SCC, whereas WR, MaxSlope, and Kep were significantly lower, indicating early enhancement and a faster dispersion model in IP-SCC. MVD was positively correlated with WR and Kep and negatively correlated with Tpeak. Tpeak was slightly positively correlated to claudin-5 expression. CONCLUSION: DCE-MRI can serve as a noninvasive biomarker of angiogenesis in the malignant transformation from SIP to IP-SCC. DCE-MRI may assist in the differentiation of malignancies and treatment selection.

Investigation of the Value of T 2 Mapping in the Prediction of Eosinophilic Chronic Rhinosinusitis With Nasal Polyps.

OBJECTIVES: Patients with eosinophilic chronic rhinosinusitis with nasal polyps (eosCRSwNP) usually have more extensive sinus disease, severe symptoms, and poorer disease control compared with patients with non-eosCRSwNP. Separating these entities will be crucial for patient management. The purpose of this study is to investigate T 1, T 2 , and apparent diffusion coefficient (ADC) values of the nasal polyps in patients with CRSwNP and evaluate the usefulness of these parameters for differentiating these diseases. METHODS: Sinonasal magnetic resonance imaging was performed in 36 patients with eosCRSwNP and 20 patients with non-eosCRSwNP (including T 1 mapping, T 2 mapping, and diffusion-weighted imaging) before surgery. The T 1 , T 2 , and ADC values were calculated and correlated with pathologically assessed inflammatory cells of nasal polyps. RESULTS: Significant higher T 2 value, higher eosinophil count, and lower lymphocyte count of the nasal polyps were observed in eosCRSwNP than those in non-eosCRSwNP. There was no significant difference in T 1 or ADC values between the 2 groups. T 2 value was correlated with eosinophil count and lymphocyte count in CRSwNP. The area under the curve of T 2 value for predicting eosCRSwNP was 0.78 with 89.9% sensitivity and 60.0% specificity. CONCLUSION: T 2 value is a promising imaging biomarker for predicting eosCRSwNP. It can help to distinguish eosCRSwNP from non-eosCRSwNP.

Dynamic contrast-enhanced MRI can quantitatively identify malignant transformation of sinonasal inverted papilloma.

OBJECTIVE: To investigate the diagnostic performance of quantitative and semi-quantitative parameters derived from dynamic contrast-enhanced MRI (DCE-MRI) in differentiating sinonasal inverted papilloma (SIP) from SIP with coexisting malignant transformation into squamous cell carcinoma (MT-SIP). METHODS: This retrospective study included 122 patients with 88 SIP and 34 MT-SIP. Quantitative and semi-quantitative parameters derived from DCE-MRI were compared between SIP and MT-SIP. The multivariate logistic regression analysis was performed to identify independent indicators and construct regression model for distinguishing MT-SIP and SIP. Diagnostic performance of independent indicators and regression model were evaluated using receiver operating coefficient (ROC) analysis and compared using DeLong test. RESULTS: There were significant differences in maximum slope of increase, contrast-enhancement ratio, bolus arrival time, volume of extravascular extracellular space (Ve), and rate constant (Kep) between SIP and MT-SIP (p < 0.05). There were no significant differences in initial area under the gadolinium curve (p = 0.174) and volume transfer constant (p = 0.105) between two groups. Multivariate analysis results showed that Ve and Kep were identified as the independent indicators for differentiating MT-SIP from SIP (p < 0.001). Areas under the ROC curves (AUCs) for predicting MT-SIP were 0.779 for Ve and 0.766 for Kep. The AUC of the combination of Ve and Kep was 0.831, yielding 83% specificity and 76.5% sensitivity. CONCLUSION: DCE-MRI can quantitatively differentiate between MT-SIP and SIP. The combination of Ve and Kep yielded an optimal performance for discriminating SIP from its malignant mimics. ADVANCES IN KNOWLEDGE: DCE-MRI with quantitative and semi-quantitative parameters can provide valuable evidences for quantitatively identifying MT-SIP.

Budesonide repairs decreased barrier integrity of eosinophilic nasal polyp epithelial cells caused by PM2.5.

BACKGROUND: Eosinophilic chronic rhinitis with nasal polyps (eos-CRSwNP) is a subtype of nasal polyps (NPs) characterized by severe type-2 inflammation and defective epithelial barrier function. The epithelial barrier plays important roles in the pathogenesis of NPs and type-2 inflammation. Particular matter 2.5 (PM2.5) are fine particles with a diameter less than 2.5 µm, containing a mixture of different components. Here, we investigated the impact of PM2.5 on the barrier function of the eos-CRSwNP epithelium and explored the reparative function of budesonide. METHODS: Samples from noninflammatory nasal mucosa and eos-CRSwNP were collected to establish an in vitro air-liquid interface cultured model. The cells were exposed to PM2.5 at 50 or 100 µg/ml intermittently for 72 h, with or without budesonide pretreatment. Barrier function and tight junction (TJ) expression were reflected by measuring transepithelial resistance (TER), paracellular flux permeability of fluorescein isothiocyanate-labeled 4-kDa dextran, quantitative real-time polymerase chain reaction (qPCR), and immunofluorescence staining of TJ proteins. Cytokine expression was measured by qPCR and enzyme-linked immunosorbent assay or Luminex. RESULTS: PM2.5 increased paracellular flux and downregulated TJ protein expression (zona occuldens-1, occludin, and claudin-1), but did not change TER. These changes could be partially restored by budesonide treatment. Interleukin (IL)-8, IL-10, IL-1α, and tissue inhibitor of metalloproteinase (TIMP)-1 concentrations were significantly increased in the culture medium of cells exposed to PM2.5, and budesonide significantly reduced the changes in IL-8, IL-1α, and TIMP-1. CONCLUSION: PM2.5 impaired the barrier function of eos-CRSwNP epithelial cells and increased the permeability of large molecules. PM2.5 also increased the secretion of pro-inflammatory cytokines by nasal epithelial cells. Budesonide could partially repair the damage, suggesting potential applications in clinical practice.

Changes in Clinical and Histological Characteristics of Nasal Polyps in Northern China over the Past 2-3 Decades.

INTRODUCTION: Recent studies have shown that inflammatory patterns of nasal polyps from patients with chronic rhinosinusitis (CRS) with nasal polyps (CRSwNP) in East Asia have changed over time. However, to date there is a marked lack of similar data for CRSwNP in Northern China. This study thus aimed to assess the changes in the clinical and histological characteristics of CRSwNP patients from Northern China over the past 2-3 decades. METHODS: This was a retrospective study, which examined data from 2 groups of 150 CRSwNP patients each, who had undergone endoscopic sinus surgery in Beijing Tongren Hospital from 1993 to 1995 (group A) and from 2015 to 2019 (group B). All relevant data for demographic, clinical, and histological parameters were collected for each patient from the 2 groups and compared for overall changes between the 2 groups. RESULTS: The comorbidity of CRSwNP and asthma increased over time and the cellular phenotype of CRSwNPchanged significantly; in particular, the proportion of eosinophil-dominant CRSwNP increased, lymphocyte-dominant and plasma-dominant CRSwNP decreased significantly, and the proportions of neutrophil-dominant and mixed CRSwNP were not altered. The rate of polyp recurrence increased in CRSwNP but did not in eosinophilic CRSwNP. Smoking and age did not significantly impact the inflammatory patterns of CRSwNP. CONCLUSIONS: The inflammatory patterns of CRSwNP patients have changed and comorbidity of asthma significantly increased in CRSwNP patients in Northern China over the past 2-3 decades.

Dynamic Contrast-Enhanced MRI Can Quantitatively Discriminate the Original Site From Peripheral Portion of Sinonasal Inverted Papillomas.

BACKGROUND: Identification of the original site of sinonasal inverted papillomas (SIPs) is difficult but essential for reducing the recurrence rate. Dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) may provide information about tissue perfusion and permeability to solve this problem. PURPOSE: To investigate the accuracy of DCE-MRI parameters in discriminating between regions of interest (ROIs) in the original site and peripheral portion. STUDY TYPE: Retrospective. POPULATION: Ninety consecutive patients with pathologically proven SIP. FIELD STRENGTH/SEQUENCE: 3.0T/DCE-MRI using fast-spoiled gradient recalled (FSPGR) T1 -weighted images with fat saturation. ASSESSMENT: ROIs were placed in the original site and the peripheral portion of SIP by two radiologists according to surgical records. Maximum slope of increase (MaxSlope), contrast-enhancement ratio (CER), bolus arrival time (BAT), initial area under the signal intensity-time curve (IAUGC), volume transfer constant (Ktrans ), volume of the extravascular extracellular space (ve ), and rate constant (Kep ) were calculated and repeated again with a month interval by a radiologist. STATISTICAL TESTS: Univariate and multivariate analysis was used to determine the best diagnostic parameters, and their performances in discrimination were evaluated by receiver operating characteristic (ROC) curves. Reproducibility was estimated by the intraclass correlation coefficient (ICC). RESULTS: MaxSlope, CER, IAUGC, Ktrans , and ve were significantly lower (P < 0.05) in the original site than the peripheral portion of SIPs. CER (odds ratio [OR] = 0.227, 95% confidence interval [95% CI] = 0.073-0.704) and ve (OR = 0.048, 95% CI = 0.004-0.527) were the best indicators for identifying the original ROIs. The combination of CER and ve had the best diagnostic performance in the discrimination between the ROIs (the area under the curve [AUC]: 0.937; 95% CI: 0.896-0.974). DATA CONCLUSION: DCE-MRI derived parameter values differed between the original site and the peripheral portion of SIPs. The model combining CER and ve appears to be able to accurately distinguish the original from peripheral ROIs. LEVEL OF EVIDENCE: 4 TECHNICAL EFFICACY STAGE: 2.

Particulate Matter 2.5 Causes Deficiency in Barrier Integrity in Human Nasal Epithelial Cells.

PURPOSE: The effect of air pollution-related particulate matter (PM) on epithelial barrier function and tight junction (TJ) expression in human nasal mucosa has not been studied to date. This study therefore aimed to assess the direct impact of PM with an aerodynamic diameter less than 2.5 µ (PM2.5) on the barrier function and TJ molecular expression of human nasal epithelial cells. METHODS: Air-liquid interface cultures were established with epithelial cells derived from noninflammatory nasal mucosal tissue collected from patients undergoing paranasal sinus surgery. Confluent cultures were exposed to 50 or 100 µg/mL PM2.5 for up to 72 hours, and assessed for 1) epithelial barrier integrity as measured by transepithelial resistance (TER) and permeability of fluorescein isothiocyanate (FITC) 4 kDa; 2) expression of TJs using real-time quantitative polymerase chain reaction and immunofluorescence staining, and 3) proinflammatory cytokines by luminometric bead array or enzyme-linked immunosorbent assay. RESULTS: Compared to control medium, 50 and/or 100 µg/mL PM2.5-treatment 1) significantly decreased TER and increased FITC permeability, which could not be restored by budesonide pretreatment; 2) significantly decreased the expression of claudin-1 messenger RNA, claudin-1, occludin and ZO-1 protein; and 3) significantly increased production of the cytokines interleukin-8, TIMP metallopeptidase inhibitor 1 and thymic stromal lymphopoietin. CONCLUSIONS: Exposure to PM2.5 may lead to loss of barrier function in human nasal epithelium through decreased expression of TJ proteins and increased release of proinflammatory cytokines. These results suggest an important mechanism of susceptibility to rhinitis and rhinosinusitis in highly PM2.5-polluted areas.

[The clinic and pathologic significance of plasma cell myeloma with CCND1].

OBJECTIVE: To study the clinical and pathologic features of multiple myeloma(MM) with CCND1. METHODS: Retrospectively analyzed the clinical and pathologic profiles of 158 patients with MM from 2010 to 2013. The clinical and morphologic features of bone marrow aspiration, biopsy and immunophenotypic analysis which was carried out by flow cytometry and immunohistochemistry were analyzed in all patients with MM respectively. CCND1 translocation was studied by FISH method in all cases. Classical cytogenetic studies of bone marrow were performed in 24 cases whose CCND1 was positive. RESULTS: In the 158 patients with MM, CCND1 was detected in 31 patients (19.6%）. In 31 patients, type IgA, IgD, IgG, IgM, light-chain only and nonsecretory MM were 4 cases,4 cases,11 cases,1 case, 6 cases and 5 cases respectively. A high incidence of CCND1 was observed in IgD and nonsecretory MM comparied with IgA and IgG respectively (P<0.05). but no statistical significance was reached between κ and λ type patients (P=0.627). The morphology of plasma cell in bone marrow biopsies were small Lymphocyte- Like 24 cases,mature plasma cell 6 cases and immature plasma cell 1 case. Immunophenotype of all 31 cases was CD38⁺CD138⁺CD19⁻CD45⁻, (CD56⁺ in 11 cases, CD20⁺ in 9 cases, CD117⁺ in 3 cases. MM with CCND1 showed a strong association with CD20 expression, the lack of CD56 expression. Immunohistochemistry showed positive for cyclinD1 in 22 cases. CONCLUSION: A high incidence of CCND1 was detected in the IgD and nonsecretory MM, and correlated with Small Lymphocyte- Like, higher positive rate of CD20, cyclinD1 and the lack of CD56 expression. MM with CCND1 must be distinguished from LPL and other mature B cell lymphomas which have plasmacytoid differentiation.

[The role of intracellular Ca2+ release in olfactory signal transduction].

OBJECTIVE: To setup the real time monitor system of the concentration of free intracellular calcium ([Ca2+]i) of olfactory receptor neurons (ORNs) cultured from olfactory epithelium explant, and to analyze the role of several important components in olfactory signal transduction. METHOD: The [Ca2+]i of the cultured ORNs was determined by fluorescence microscopy using the fluorescent calcium indicator, Fura-2 AM, and calculated by means of dual-wavelength ratiometric method. Forskolin and IBMX were used to stimulate the cultured ORNs respectively. The source of corresponding [Ca2+]i elevation was studied by the depletion of extracellular or intracellular calcium. RESULT: The [Ca2+]i of silent ORNs was (58.5 +/- 12.8) nmol/L. Forskolin or IBMX stimulation led to reversible accumulation of [Ca2+]i in the ORNs. The [Ca2+]i change was abolished with the removal of extracellular Ca2+ and un-affected by treatment with thapsigargin. CONCLUSION: A system to visualize and quantify [Ca2+]i of the ORNs was established. [Ca2+]i of the ORNs was regulated by second messenger gated calcium channels.

Olfactory event-related potential in patients with rhinosinusitis-induced olfactory dysfunction.

BACKGROUND: Olfactory disorders are a common syndrome in the rhinology clinic. Olfactory event-related potential (OERP) has been considered as an important alternative method to evaluate olfactory function. The aim of this study was to find the consistency of OERP and the psychophysics olfactory test (T&T olfactometry assessment) in rhinosinusitis patients with or without nasal polyposis. METHODS: Both the psychophysics olfactory test and the OERP were performed in all patients before functional endoscopic sinus surgery (FESS). Detailed information including demographic, comorbidity, subjective symptoms, CT, and endoscopic examination were recorded. Six months later, the T&T test and OERP were reexamined in patients with preoperative identified OERP. RESULTS: Preoperative OERP was identified in 26 patients of the nonpolyp group and 12 patients of the polyp group. Good correlation existed between T&T scores and amplitude and latency of N1 and P2 in 26 nonpolyp patients (p < 0.05). No meaningful correlation was apparent in 12 polyp patients (p > 0.05). Significant correlation existed in 7 polyp patients with a free olfactory cleft (OC) according to CT (p < 0.05). The other 5 patients with partial OC obstruction were anosmic according to the psychophysics olfactory test; however, OERP was present. Six months later, T&T scores decreased significantly only in these five patients (p < 0.05). Significant correlation was obtained between postoperative T&T scores and OERP in both polyp and nonpolyp groups (p < 0.05). CONCLUSION: OERP can be used to investigate olfactory function of rhinosinusitis patients. Nasal polyp obstruction in specific parts of the OC might influence the consistency between the psychophysics olfactory test and OERP.

Effects of transplanting olfactory ensheathing cells on recovery of olfactory epithelium after olfactory nerve transection in rats.

BACKGROUND: Olfactory ensheathing cells (OECs) can remyelinate injured spinal cord and the peripheral nerve system, but little is known about its effect on the transected olfactory nerve. We investigated recovery of olfactory epithelium after transplanting allogeneic OECs in transected rat olfactory nerves. MATERIAL/METHODS: Olfactory ensheathing cells from the olfactory bulb were cultured in DMEM/F-12 medium and purified with cytosine arabinoside (Ara-c). Forty Sprague-Dawley rats were divided into 2 groups; the left olfactory nerve was transected in all animals. In the transplant group, DiI-labeled OECs were injected into the gap between the dura and the cribriform plate (n=24); DMEM/F-12 medium was injected in control animals (n=16). Rats were subsequently killed for histologic examination. Olfactory evoked potentials (OEPs) were used to evaluate nerve conduction. RESULTS: After transecting the olfactory nerve, there was no horseradish peroxidase staining in the olfactory bulb; some OEPs disappeared. Five days after surgery, there was no horseradish peroxidase staining in the olfactory bulb of any animal. Apoptotic cells appeared in the epithelium; the thickness and cell number of the olfactory epithelium were decreased. Two weeks later, the thickness and cell number of the olfactory epithelium increased gradually. Some horseradish peroxidase staining in the olfactory bulb and OECs was detected; more growth associated protein-43 marked olfactory receptor neurons were visible. Six weeks after surgery, the cell number was greater in the transplant group (P<0.05); there was no statistically significant between-group difference regarding olfactory epithelium thickness. CONCLUSIONS: Transplanted OECs may be used to treat transected olfactory nerves.

[Impact of olfactory nerve transection on the apoptosis of mice olfactory receptor neurons].

OBJECTIVE: To analyze the impact of olfactory nerve transection on the apoptosis of mice olfactory receptor neurons (ORN), and discuss the reliability of this experimental model. METHODS: After olfactory nerve transection of mice, anterograde horseradish peroxidase (HRP) tracing was performed to confirm the completion of nerve transection. On 8 h, 2 d, 3 d and 5 d after surgery, TdT mediated deoxyuridine triphosphate-biotin nick end labelling (TUNEL) was used to observe the apoptosis of ORN, while relative semi-quantitative RT-PCR and immunohistochemistry were used to reflect the expression of olfactory marker protein (OMP, special marker of mature ORN) in olfactory epithelium. RESULTS: No HRP label was observed in olfactory bulb after olfactory nerve transaction. Both TUNEL-positive and OMP-positive cells were ORN. After the surgery, TUNEL-positive cells increased remarkably and peaked on 2 d after the surgery. Meanwhile OMP mRNA in olfactory epithelium began to decrease markedly till 5 d after the surgery, and the olfactory epithelium got thinner accordingly. CONCLUSIONS: This experimental model can be used reliably to sever mice olfactory nerve and manipulate simultaneous apoptosis of mice ORN.