Targeting CD13/aminopeptidase N as a novel therapeutic approach for scleroderma fibrosis.

OBJECTIVE: Systemic sclerosis (SSc) is an autoimmune multisystem disease with poorly understood pathogenesis and ineffective treatment options. Soluble CD13 (sCD13), generated by cleavage of cell surface CD13 via matrix metalloproteinase 14 (MMP14), signals through the bradykinin receptor B1 (B1R) to elicit pro-inflammatory, pro-arthritic, and pro-angiogenic responses. In this study we explored the anti-fibrotic potential of targeting the sCD13-B1R axis in SSc. METHODS: The expression of CD13, B1R and MMP14 was examined in SSc skin and explanted dermal fibroblasts. The efficacy of B1R antagonists in the inhibition on fibrosis was determined in vitro and in vivo. RESULTS: Expression of the genes for CD13, B1R and MMP14 was elevated in skin biopsies from patients with diffuse cutaneous (dc)SSc. Notably, single cell analysis of SSc skin biopsies revealed the highest BDKRB1 expression in COL8A1-positive myofibroblasts, a population exclusively seen in SSc. TGF-ß induced the expression of BDKRB1 and production of sCD13 by dcSSc skin fibroblasts. Treatment of dcSSc fibroblasts with sCD13 promoted fibrotic gene expression, signaling, cell proliferation, migration, and gel contraction. The profibrotic sCD13 or TGFß responses were prevented by a B1R antagonist. Mice lacking Cd13 or Bdkrb1 were resistant to bleomycin-induced skin fibrosis and inflammation. Pharmacological B1R inhibition had a comparable antifibrotic effect. CONCLUSION: These results are the first to demonstrate a key role for sCD13 in SSc skin fibrosis, and suggest that targeting the sCD13-B1R signaling axis is a promising novel therapeutic approach for SSc.

Tobacco smoking and nicotine vaping in persons with first episode psychosis.

Tobacco smoking is highly prevalent in persons with psychosis and is the leading cause of preventable mortality in this population. Less is known about tobacco smoking in persons with first episode psychosis (FEP) and there have been no estimates about the prevalence of nicotine vaping in FEP. This study reports rates of tobacco smoking and nicotine vaping in young people with FEP enrolled in Coordinated Specialty Care programs in Pennsylvania and Maryland. Using data collected from 2021 to 2023, we examined lifetime and recent smoking and vaping and compared smokers and vapers to nonusers on symptoms, functioning, and substance use. The sample included 445 participants aged 13-35 with recent psychosis onset. Assessments were collected by program staff. Overall, 28 % of participants engaged in either smoking or vaping within 30 days of the admission assessment. Smokers and vapers were disproportionately male, cannabis users, and had lower negative symptom severity than non-smokers. Vapers had higher role and social functioning. Both smoking and vaping were related to a longer time from psychosis onset to program enrollment. We compare these findings to previous studies and suggest steps for addressing smoking and vaping in this vulnerable population.

Activation of cytotoxic lymphocytes through CD6 enhances killing of cancer cells.

Immune checkpoint inhibitors (ICIs) have demonstrated efficacy and improved survival in a growing number of cancers. Despite their success, ICIs are associated with immune-related adverse events that can interfere with their use. Therefore, safer approaches are needed. CD6, expressed by T-lymphocytes and human NK cells, engages in cell-cell interactions by binding to its ligands CD166 (ALCAM) and CD318 (CDCP1). CD6 is a target protein for regulating immune responses and is required for the development of several mouse models of autoimmunity. Interestingly, CD6 is exclusively expressed on immune cells while CD318 is strongly expressed on most cancers. Here we demonstrate that disrupting the CD6-CD318 axis with UMCD6, an anti-CD6 monoclonal antibody, prolongs survival of mice in xenograft mouse models of human breast and prostate cancer, treated with infusions of human lymphocytes. Analysis of tumor-infiltrating immune cells showed that augmentation of lymphocyte cytotoxicity by UMCD6 is due to effects of this antibody on NK, NKT and CD8 + T cells. In particular, tumor-infiltrating cytotoxic lymphocytes from UMCD6-treated mice expressed higher levels of perforin and were found in higher proportions than those from IgG-treated mice. Moreover, RNA-seq analysis of human NK-92 cells treated with UMCD6 revealed that UMCD6 up-regulates the NKG2D-DAP10 receptor complex, important in NK cell activation, as well as its downstream target PI3K. Our results now describe the phenotypic changes that occur on immune cells upon treatment with UMCD6 and further confirm that the CD6-CD318 axis can regulate the activation state of cytotoxic lymphocytes and their positioning within the tumor microenvironment.

Activation of Cytotoxic Lymphocytes Through CD6 Enhances Killing of Cancer Cells.

Immune checkpoint inhibitors (ICIs) have demonstrated efficacy and improved survival in a growing number of cancers. Despite their success, ICIs are associated with immune-related adverse events that can interfere with their use. Therefore, safer approaches are needed. CD6, expressed by T-lymphocytes and human NK cells, engages in cell-cell interactions by binding to its ligands CD166 (ALCAM) and CD318 (CDCP1). CD6 is a target protein for regulating immune responses and is required for the development of several mouse models of autoimmunity. Interestingly, CD6 is exclusively expressed on immune cells while CD318 is strongly expressed on most cancers. Here we demonstrate that disrupting the CD6-CD318 axis with UMCD6, an anti-CD6 monoclonal antibody, prolongs survival of mice in xenograft models of human breast and prostate cancer, treated with infusions of human lymphocytes. Analysis of tumor-infiltrating immune cells showed that augmentation of lymphocyte cytotoxicity by UMCD6 is due to effects of this antibody on NK, NKT and CD8+ T cells. Tumor-infiltrating cytotoxic lymphocytes were found in higher proportions and were activated in UMCD6-treated mice compared to controls. Similar changes in gene expression were observed by RNA-seq analysis of NK cells treated with UMCD6. Particularly, UMCD6 up-regulated the NKG2D-DAP10 complex and activated PI3K. Thus, the CD6-CD318 axis can regulate the activation state of cytotoxic lymphocytes and their positioning within the tumor microenvironment.

Mammalian Glycosylation Patterns Protect Citrullinated Chemokine MCP-1/CCL2 from Partial Degradation.

Monocyte chemoattractant protein-1 (MCP-1/CCL2) is a potent chemotactic agent for monocytes, primarily produced by macrophages and endothelial cells. Significantly elevated levels of MCP-1/CCL2 were found in synovial fluids of patients with rheumatoid arthritis (RA), compared to osteoarthritis or other arthritis patients. Several studies suggested an important role for MCP-1 in the massive inflammation at the damaged joint, in part due to its chemotactic and angiogenic effects. It is a known fact that the post-translational modifications (PTMs) of proteins have a significant impact on their properties. In mammals, arginine residues within proteins can be converted into citrulline by peptidylarginine deiminase (PAD) enzymes. Anti-citrullinated protein antibodies (ACPA), recognizing these PTMs, have become a hallmark for rheumatoid arthritis (RA) and other autoimmune diseases and are important in diagnostics and prognosis. In previous studies, we found that citrullination converts the neutrophil attracting chemokine neutrophil-activating peptide 78 (ENA-78) into a potent macrophage chemoattractant. Here we report that both commercially available and recombinant bacterially produced MCP-1/CCL2 are rapidly (partially) degraded upon in vitro citrullination. However, properly glycosylated MCP-1/CCL2 produced by mammalian cells is protected against degradation during efficient citrullination. Site-directed mutagenesis of the potential glycosylation site at the asparagine-14 residue within human MCP-1 revealed lower expression levels in mammalian expression systems. The glycosylation-mediated recombinant chemokine stabilization allows the production of citrullinated MCP-1/CCL2, which can be effectively used to calibrate crucial assays, such as modified ELISAs.

Soluble CD13 induces inflammatory arthritis by activating the bradykinin receptor B1.

CD13, an ectoenzyme on myeloid and stromal cells, also circulates as a shed, soluble protein (sCD13) with powerful chemoattractant, angiogenic, and arthritogenic properties, which require engagement of a G protein-coupled receptor (GPCR). Here we identify the GPCR that mediates sCD13 arthritogenic actions as the bradykinin receptor B1 (B1R). Immunofluorescence and immunoblotting verified high expression of B1R in rheumatoid arthritis (RA) synovial tissue and fibroblast-like synoviocytes (FLSs), and demonstrated binding of sCD13 to B1R. Chemotaxis, and phosphorylation of Erk1/2, induced by sCD13, were inhibited by B1R antagonists. In ex vivo RA synovial tissue organ cultures, a B1R antagonist reduced secretion of inflammatory cytokines. Several mouse arthritis models, including serum transfer, antigen-induced, and local innate immune stimulation arthritis models, were attenuated in Cd13-/- and B1R-/- mice and were alleviated by B1R antagonism. These results establish a CD13/B1R axis in the pathogenesis of inflammatory arthritis and identify B1R as a compelling therapeutic target in RA and potentially other inflammatory diseases.

Inhibition of bromodomain extraterminal histone readers alleviates skin fibrosis in experimental models of scleroderma.

Binding of the bromodomain and extraterminal domain proteins (BETs) to acetylated histone residues is critical for gene transcription. We sought to determine the antifibrotic efficacy and potential mechanisms of BET inhibition in systemic sclerosis (SSc). Blockade of BETs was done using a pan-BET inhibitor, JQ1; BRD2 inhibitor, BIC1; or BRD4 inhibitors AZD5153 or ARV825. BET inhibition, specifically BRD4 blockade, showed antifibrotic effects in an animal model of SSc and in patient-derived diffuse cutaneous SSc (dcSSc) fibroblasts. Transcriptome analysis of JQ1-treated dcSSc fibroblasts revealed differentially expressed genes related to extracellular matrix, cell cycle, and calcium (Ca2+) signaling. The antifibrotic effect of BRD4 inhibition was mediated at least in part by downregulation of Ca2+/calmodulin-dependent protein kinase II α and reduction of intracellular Ca2+ concentrations. On the basis of these results, we propose targeting Ca2+ pathways or BRD4 as potentially novel therapeutic approaches for progressive tissue fibrosis.

Effects of placebo administration on immune mechanisms and relationships with central endogenous opioid neurotransmission.

Behavioral conditioning and expectation can have profound impact on animal and human physiology. Placebo, administered under positive expectation in clinical trials, can have potent effects on disease pathology, obscuring active medications. Emerging evidence suggests placebo-responsive neurotransmitter systems (e.g., endogenous opioid) regulate immune function by manipulating inflammatory proteins including IL-18, a potent pro-inflammatory, nociceptive cytokine implicated in pathophysiology of various diseases. Validation that neuroimmune interactions involving brain µ-opioid receptor (MOR) activity and plasma IL-18 underlie placebo analgesic expectation could have widespread clinical applications. Unfortunately, current lack of mechanistic clarity obfuscates clinical translation. To elucidate neuroimmune interactions underlying placebo analgesia, we exposed 37 healthy human volunteers to a standardized pain challenge on each of 2 days within a Positron Emission Tomography (PET) neuroimaging paradigm using the MOR selective radiotracer, 11C-Carfentanil (CFN). Each day volunteers received an intervention (placebo under analgesic expectation or no treatment), completed PET scanning, and rated their pain experience. MOR BPND parametric maps were generated from PET scans using standard methods. Results showed placebo reduced plasma IL-18 during pain (W74 = -3.7, p < 0.001), the extent correlating with reduction in pain scores. Placebo reduction in IL-18 covaried with placebo-induced endogenous opioid release in the left nucleus accumbens (T148 = 3.33; puncorr < 0.001) and left amygdala (T148 = 3.30; puncorr < 0.001). These findings are consistent with a modulating effect of placebo (under analgesic expectation in humans) on a potent nociceptive, pro-inflammatory cytokine (IL-18) and underlying relationships with endogenous opioid activity, a neurotransmitter system critically involved in pain, stress, and mood regulation.

Short-term skeletal and dentoalveolar effects of overexpansion.

OBJECTIVES: To evaluate whether the amount of rapid maxillary expansion differentially affects the skeletal and dentoalveolar changes that occur. MATERIALS AND METHODS: This randomized controlled trial included 23 patients who had rapid maxillary expansion (RME). Subjects were randomly assigned to a conventional expansion control group (n = 12) or an overexpansion group (n = 11), who started treatment at 13.2 ± 1.5 and 13.8 ± 1 years of age, respectively. Cone beam computed tomography scans (11 cm) were obtained prior to rapid maxillary expander (RME) delivery and approximately 3.7 months later. Initial hand-wrist radiographs were used to determine the participants' skeletal maturity. RESULTS: The RME screws were activated 5.6 ± 1.2 mm and 10.1 ± 0.6 mm in the conventional and overexpansion groups, respectively. Overexpansion produced significantly greater expansion of the nasal cavity (2.1X-2.5X), maxillary base (2.3X), buccal alveolar crest (1.4X), and greater palatine foramina (1.9X). Significantly greater intermolar width increases (1.8X) and molar inclination (2.8X) changes were also produced. The nasal cavity and maxillary base expanded 23%-32% as much as the screws were activated. Skeletal expansion was positively correlated with RME screw activation (R = 0.61 to 0.70) and negatively correlated (R = -0.56 to -0.64) with the patients' skeletal maturation indicators (SMIs). Together, screw activation and the patients' SMI scores explained 48%-66% of the variation in skeletal expansion. CONCLUSIONS: This pilot study shows that overexpansion produces greater changes than conventional expansion, with greater skeletal effects among less mature patients.

How different time intervals between repeated applications of CPP-ACP fluoride varnish effect smooth surface enamel demineralization?

OBJECTIVES: To evaluate the preventive effects of different time intervals between repeated applications of the CPP-ACP fluoride varnish on enamel demineralization. METHODS: Human teeth were sectioned and randomly allocated to three groups: 4-week, 6-week, and 12-week (N = 22/group). Baseline images of the enamel surfaces were obtained using the FluoreCam recording the area, intensity, and impact of baseline enamel demineralization. All groups received fluoride varnish applications at the beginning of the experiment. The varnish was reapplied every 4 or 6 weeks in the 4-week and 6-week groups, respectively. Following each application, the groups underwent thermo-cycling, tooth brushing and pH cycling to simulate the time effect. After 12 weeks, the enamel surfaces were reimaged using the FluoreCam. Within and between-group differences in the area, intensity and impact of demineralization were evaluated. RESULTS: At baseline, there were no significant between-group differences for area, intensity, or impact. Statistically significant (p<0.001) enamel demineralization occurred over time within each group. There were significant between-group differences in the changes that occurred in area (P = 0.004), impact (P = 0.022), but not intensity. The 12-week had significantly larger areas of demineralization than the 6-week (P = 0.041) and 4-week (P = 0.001) groups. Changes in impact was significantly (P = 0.007) greater in the 12-week group than 4-week group, but not greater than the 6-week group. There were no statistically significant differences between 4- and 6-week groups in the changes of area, intensity, or impact. CONCLUSION: Reapplication of the CPP-ACP fluoride varnish every 4-6 weeks, is more effective in reducing enamel demineralization compared to every 12 weeks.

Stability of immediately loaded 3 mm long miniscrew implants: a feasibility study.

INTRODUCTION: Shorter miniscrew implants (MSIs) are needed to make orthodontics more effective and efficient. OBJECTIVE: To evaluate the stability, insertion torque, removal torque and pain associated with 3 mm long MSIs placed in humans by a novice clinician. METHODS: 82 MSIs were placed in the buccal maxillae of 26 adults. Pairs of adjacent implants were immediately loaded with 100g. Subjects were recalled after 1, 3, 5, and 8 weeks to verify stability and complete questionnaires pertaining to MSI-related pain and discomfort. RESULTS: The overall failure rate was 32.9%. The anterior and posterior MSIs failed 35.7% and 30.0% of the time, respectively. Excluding the 10 MSIs (12.2%) that were traumatically dislodged, the failure rates in the anterior and posterior sites were 30.1% and 15.2%, respectively; the overall primary failure rate was 23.6%. Failures were significantly (p= 0.010) greater (46.3% vs 19.5%) among the first 41 MSIs than the last 41 MSIs that were placed. Excluding the traumatically lost MSIs, the failures occurred on or before day 42. Subjects experienced very low pain (2.2% of maximum) and discomfort (5.5% of maximum) during the first week only. CONCLUSIONS: Shorter 3 mm MSIs placed by a novice operator are highly likely to fail. However, failure rates can be substantially decreased over time with the placement of more MSIs. Pain and discomfort experienced after placing 3 mm MSIs is minimal and temporary.

The quality of etched enamel in different regions and tooth types and its significance in bonding and the development of white spot lesions.

OBJECTIVES: To quantify differences in the etch quality of enamel within and between human teeth, which has not previously been attempted. MATERIALS AND METHODS: The buccal right and left halves of 27 extracted human teeth were randomly allocated to scanning electron microscopy (SEM) or micro-computed tomography (µCT) for evaluation. The buccal surfaces were pumiced, etched with 37% phosphoric acid gel etchant for 15 seconds, rinsed, and air dried. Each tooth was divided into three regions (incisal, middle, and cervical) and viewed after etching at 1200× magnification with SEM. The µCT scans were taken before and after etching to calculate apparent and material mineral densities. RESULTS: SEM showed greater aprismatic enamel and poorer etch quality (ie, significantly less percentage enamel) for the posterior than anterior teeth and for the cervical region than for the incisal and middle regions of all teeth. Although there were no density differences prior to etching, µCT demonstrated that etching increased material density significantly more for the anterior than posterior teeth. Prior to etching, the enamel in the cervical regions was significantly less dense than the enamel in the middle or incisal regions. Etching significantly increased the material density of all three regions, which decreased initial regional differences. After etching, the apparent density of the cervical region remained significantly lower than the densities of the other two regions. CONCLUSIONS: Based on SEM and µCT, there is greater aprismatic enamel and inferior etch quality in the cervical regions of all tooth types and is clinically significant in explaining the failure of sealant retention and the propensity for white spot lesions.

Keys to Class II correction: A comparison of 2 extraction protocols.

INTRODUCTION: The purpose of this study was to evaluate the effects of 2 extraction patterns on incisor and molar movements in patients with growing Class II Division 1. METHODS: The sample included 54 patients 10-17 years of age treated by 2 private practice orthodontists using Tweed directional force mechanics, 4 premolar extractions, J-hook headgears, and Class II elastics or Saif springs. The sample was divided on the basis of having maxillary and mandibular first premolars (4/4) or maxillary first and mandibular second premolars (4/5) extracted. Each group included 27 patients. Treatment lasted 2.8 ± 0.60 years and 2.6 ± 0.54 years for the 4/4 and 4/5 groups, respectively. Pretreatment (T1) and posttreatment lateral cephalograms and dental casts were evaluated. Cranial base, mandibular, and maxillary superimpositions were performed to quantify tooth movements and displacements. RESULTS: There were no statistically significant T1 between-group differences in crowding or in the SNA, SNB, ANB, and MPA angles. Analyses of covariance, controlling for statistically significant (P <0.05) differences in T1 mandibular incisor position, showed that mandibular first premolars extractions produced greater (1.6 mm) mandibular incisor retraction than second premolar extractions. The mandibular first molars were protracted significantly more (0.7 mm) after the second premolar than the first premolar extractions. Within-group changes of the MPA, between-group differences in the changes in MPA, and the amount of vertical eruption of the maxillary and mandibular molars were not significantly different between the 2 extraction patterns. CONCLUSIONS: Extraction of mandibular second premolars enhances Class II molar correction, with greater mesial first molar movement and less distal incisor movement. Neither extraction pattern has an effect on the MPA or the vertical dimension (ie, there was no "wedge effect").

Lymphocyte subset abnormalities in early diffuse cutaneous systemic sclerosis.

BACKGROUND: Abnormalities in lymphocyte surface markers and functions have been described in systemic sclerosis (SSc), but conflicting results abound, and these studies often examined patients with heterogeneous disease duration, severity, clinical phenotype, and concurrent immunosuppressive agents. We studied a clinically homogeneous group of early diffuse cutaneous SSc patients not exposed to immunosuppressive drugs who were enrolled in a clinical trial and compared their immune parameters to healthy control subjects. METHODS: Lymphocyte subsets were enumerated by multi-parameter flow cytometry of peripheral blood mononuclear cells at baseline visit. Production of the cytokines IL-4 and IL-17 was measured by intracellular flow cytometry following T cell activation. RESULTS: SSc patients had increased percentages of CD4+ T cells but lower percentages of CD8+ T cells versus controls. The CD28-negative population was expanded in SSc, in the CD4 subset. Striking expansion of CD319+ T cells was noted among the CD4+ cells, in which they were barely detectable in healthy subjects. Frequencies of IL-4 producing cells did not differ between SSc and controls, but expansion of IL-17 producing cells was observed in SSc. A higher proportion of CD319+ cells produced cytokines, compared to other CD4+ cells. Numbers of activated T cells, regulatory T cells, and B cells were similar in SSc and control groups. Circulating follicular helper but not peripheral helper T cells were slightly expanded in SSc. CONCLUSION: In this carefully selected group of early diffuse cutaneous SSc patients, analysis of immune cell parameters has identified abnormalities that likely reflect disease pathogenesis and that are candidate biomarkers for sub-classification and targeted treatment. The CD4+CD319+ (SLAM-F7+) cells are cytotoxic and oligoclonal, were recently shown to be a dominant T cell population in perivascular lymphocytic infiltrates in SSc skin, actively secrete cytokines, and are emerging as a target for novel treatments of SSc.

CD6 is a target for cancer immunotherapy.

Limitations of checkpoint inhibitor cancer immunotherapy include induction of autoimmune syndromes and resistance of many cancers. Since CD318, a novel CD6 ligand, is associated with the aggressiveness and metastatic potential of human cancers, we tested the effect of an anti-CD6 monoclonal antibody, UMCD6, on killing of cancer cells by human lymphocytes. UMCD6 augmented killing of breast, lung, and prostate cancer cells through direct effects on both CD8+ T cells and NK cells, increasing cancer cell death and lowering cancer cell survival in vitro more robustly than monoclonal antibody checkpoint inhibitors that interrupt the programmed cell death 1 (PD-1)/PD-1 ligand 1 (PD-L1) axis. UMCD6 also augmented in vivo killing by human peripheral blood lymphocytes of a human breast cancer line xenotransplanted into immunodeficient mice. Mechanistically, UMCD6 upregulated the expression of the activating receptor NKG2D and downregulated expression of the inhibitory receptor NKG2A on both NK cells and CD8+ T cells, with concurrent increases in perforin and granzyme B production. The combined capability of an anti-CD6 monoclonal antibody to control autoimmunity through effects on CD4+ lymphocyte differentiation while enhancing killing of cancer cells through distinct effects on CD8+ and NK cells opens a potential new approach to cancer immunotherapy that would suppress rather than instigate autoimmunity.

Stability of immediately loaded 3 mm long miniscrew implants: a feasibility study

ABSTRACT Introduction: Shorter miniscrew implants (MSIs) are needed to make orthodontics more effective and efficient. Objective: To evaluate the stability, insertion torque, removal torque and pain associated with 3 mm long MSIs placed in humans by a novice clinician. Methods: 82 MSIs were placed in the buccal maxillae of 26 adults. Pairs of adjacent implants were immediately loaded with 100g. Subjects were recalled after 1, 3, 5, and 8 weeks to verify stability and complete questionnaires pertaining to MSI-related pain and discomfort. Results: The overall failure rate was 32.9%. The anterior and posterior MSIs failed 35.7% and 30.0% of the time, respectively. Excluding the 10 MSIs (12.2%) that were traumatically dislodged, the failure rates in the anterior and posterior sites were 30.1% and 15.2%, respectively; the overall primary failure rate was 23.6%. Failures were significantly (p= 0.010) greater (46.3% vs 19.5%) among the first 41 MSIs than the last 41 MSIs that were placed. Excluding the traumatically lost MSIs, the failures occurred on or before day 42. Subjects experienced very low pain (2.2% of maximum) and discomfort (5.5% of maximum) during the first week only. Conclusions: Shorter 3 mm MSIs placed by a novice operator are highly likely to fail. However, failure rates can be substantially decreased over time with the placement of more MSIs. Pain and discomfort experienced after placing 3 mm MSIs is minimal and temporary.

RESUMO Introdução: Mini-implantes (MIs) mais curtos são necessários para uma Ortodontia mais eficiente e efetiva. Objetivo: Avaliar a estabilidade, torque de inserção e de remoção e dor associada a MIs de 3mm instalados em humanos por um ortodontista principiante. Métodos: 82 MIs foram instalados na região vestibular da arcada superior de 26 adultos. Pares de mini-implantes adjacente receberam carga imediata de 100g. Após 1, 3, 5 e 8 semanas, os pacientes foram reavaliados para verificar a estabilidade e preencher um questionário sobre a dor e o desconforto relacionados aos MIs. Resultados: A taxa geral de falhas foi de 32,9%, sendo de 35,7% para os MIs anteriores e 30% para os MIs posteriores. Excluindo os 10 MIs que foram perdidos por trauma (12,2%), a taxa de falha nas regiões anterior e posterior foram de 30,1% e 15,2%, respectivamente e ocorreram no 420 dia ou antes. A taxa geral de falha primária foi de 23,6%. A taxa de falha foi significativamente maior (p=0,010) nos primeiros 41 MIs do que nos 41 últimos (46,3% vs. 19,5%). As experiências relacionadas à dor foram baixas (2,2% máximo), assim como ao desconforto (5,5% máximo) durante a primeira semana. Conclusão: MIs de 3mm instalados por um novato são mais propensos a falhas. Porém, as taxas de falha podem diminuir substancialmente com a instalação de mais MIs com o decorrer do tempo. A dor e o desconforto após a instalação desses dispositivos são mínimos e temporários.

Tazemetostat for patients with relapsed or refractory follicular lymphoma: an open-label, single-arm, multicentre, phase 2 trial.

BACKGROUND: Activating mutations of EZH2, an epigenetic regulator, are present in approximately 20% of patients with follicular lymphoma. We investigated the activity and safety of tazemetostat, a first-in-class, oral EZH2 inhibitor, in patients with follicular lymphoma. METHODS: This study was an open-label, single-arm, phase 2 trial done at 38 clinics or hospitals in France, the UK, Australia, Canada, Poland, Italy, Ukraine, Germany, and the USA. Eligible patients were adults (≥18 years) with histologically confirmed follicular lymphoma (grade 1, 2, 3a, or 3b) that had relapsed or was refractory to two or more systemic therapies, had an Eastern Cooperative Oncology Group performance status of 0-2, and had sufficient tumour tissue for central testing of EZH2 mutation status. Patients were categorised by EZH2 status: mutant (EZH2mut) or wild-type (EZH2WT). Patients received 800 mg of tazemetostat orally twice per day in continuous 28-day cycles. The primary endpoint was objective response rate based on the 2007 International Working Group criteria for non-Hodgkin lymphoma, assessed by an independent radiology committee. Activity and safety analyses were done in patients who received one dose or more of tazemetostat. This study is registered with ClinicalTrials.gov, NCT01897571, and follow-up is ongoing. FINDINGS: Between July 9, 2015, and May 24, 2019, 99 patients (45 in the EZH2mut cohort and 54 in the EZH2WT cohort) were enrolled in the study. At data cutoff for the analysis (Aug 9, 2019), the median follow-up was 22·0 months (IQR 12·0-26·7) for the EZH2mut cohort and 35·9 months (24·9-40·5) for the EZH2WT cohort. The objective response rate was 69% (95% CI 53-82; 31 of 45 patients) in the EZH2mut cohort and 35% (23-49; 19 of 54 patients) in the EZH2WT cohort. Median duration of response was 10·9 months (95% CI 7·2-not estimable [NE]) in the EZH2mut cohort and 13·0 months (5·6-NE) in the EZH2WT cohort; median progression-free survival was 13·8 months (10·7-22·0) and 11·1 months (3·7-14·6). Among all 99 patients, treatment-related grade 3 or worse adverse events included thrombocytopenia (three [3%]), neutropenia (three [3%]), and anaemia (two [2%]). Serious treatment-related adverse events were reported in four (4%) of 99 patients. There were no treatment-related deaths. INTERPRETATION: Tazemetostat monotherapy showed clinically meaningful, durable responses and was generally well tolerated in heavily pretreated patients with relapsed or refractory follicular lymphoma. Tazemetostat is a novel treatment for patients with follicular lymphoma. FUNDING: Epizyme.

Real time visualization of cancer cell death, survival and proliferation using fluorochrome-transfected cells in an IncuCyte® imaging system.

Cancer immunotherapy is a rapidly advancing and viable approach to treating cancer along with more traditional forms of therapy. Real-time cell analysis technologies that examine the dynamic interactions between cancer cells and the cells of the immune system are becoming more important for assessment of novel therapeutics. In this report, we use the IncuCyte® imaging system to study the killing potential of various immune cells on cancer cell lines. The IncuCyte® system tracks living cells, labeled by a red fluorescent protein, and cell death, as indicated by the caspase-3/7 reagent, which generates a green fluorescent signal upon activation of apoptotic pathways. Despite the power of this approach, obtaining commercially fluorescent cancer cell lines is expensive and limited in the range of cell lines that are available. To overcome this barrier, we developed an inexpensive method using a lentiviral construct expressing nuclear localized mKate2 red fluorescent protein to stably label cancer cells. We demonstrate that this method is effective in labeling a wide variety of cell lines, allowing for analyses of different cancers as well as different cell lines of the same type of cancer.

ASCEND: Phase III, Randomized Trial of Acalabrutinib Versus Idelalisib Plus Rituximab or Bendamustine Plus Rituximab in Relapsed or Refractory Chronic Lymphocytic Leukemia.

PURPOSE: Acalabrutinib, a highly selective, potent, Bruton tyrosine kinase inhibitor, was evaluated in this global, multicenter, randomized, open-label, phase III study in patients with relapsed/refractory (R/R) chronic lymphocytic leukemia (CLL). METHODS: Eligible patients, aged ≥ 18 years with R/R CLL, were randomly assigned 1:1 centrally and stratified by del(17p) status, Eastern Cooperative Oncology Group performance status score, and number of prior lines of therapy. Patients received acalabrutinib monotherapy or investigator's choice (idelalisib plus rituximab [I-R] or bendamustine plus rituximab [B-R]). The primary end point was progression-free survival (PFS) assessed by an independent review committee (IRC) in the intent-to-treat population. Key secondary end points included IRC-assessed overall response rate, overall survival, and safety. RESULTS: From February 21, 2017, to January 17, 2018, a total of 398 patients were assessed for eligibility; 310 patients were randomly assigned to acalabrutinib monotherapy (n = 155) or investigator's choice (n = 155; I-R, n = 119; B-R, n = 36). Patients had received a median of two prior therapies (range, 1-10). After a median follow-up of 16.1 months (range, 0.03-22.4 months), median PFS was significantly longer with acalabrutinib monotherapy (PFS not reached) compared with investigator's choice (16.5 months [95% CI, 14.0 to 17.1 months]; hazard ratio, 0.31 [95% CI, 0.20 to 0.49]; P < .0001). Estimated 12-month PFS was 88% (95% CI, 81% to 92%) for acalabrutinib and 68% (95% CI, 59% to 75%) for investigator's choice. Serious adverse events occurred in 29% of patients (n = 44 of 154) treated with acalabrutinib monotherapy, 56% (n = 66 of 118) with I-R, and 26% (n = 9 of 35) with B-R. Deaths occurred in 10% (n = 15 of 154), 11% (n = 13 of 118), and 14% (n = 5 of 35) of patients receiving acalabrutinib monotherapy, I-R, and B-R, respectively. CONCLUSION: Acalabrutinib significantly improved PFS compared with I-R or B-R and has an acceptable safety profile in patients with R/R CLL.