Functional testing, coronary artery calcifications, and outcomes in Hodgkin lymphoma survivors treated with chest radiation.

BACKGROUND: Consensus guidelines recommend periodic screening for coronary artery disease (CAD) in Hodgkin lymphoma (HL) survivors treated with radiation therapy (RT) to the chest. However, the prognostic utility of screening strategies in this population remains unclear. We evaluated the association between functional testing, coronary artery calcifications (CAC), and guideline-based risk assessment and major adverse cardiovascular events (MACE) in HL survivors treated with RT. METHODS: We retrospectively studied HL survivors treated with RT who underwent functional testing between 2003 and 2020 and chest computed tomography (CT) within 12 months of each other at our center. CAC was assessed semi-quantitatively from CT images. Cardiovascular risk was estimated using the 2019 ACC/AHA Guideline on the Primary Prevention of Cardiovascular Disease. Diagnostic test characteristics were calculated using major adverse cardiac events (MACE) during follow-up as the gold standard. RESULTS: The study included 159 patients (median age at functional testing 48 years, median age at HL diagnosis 27 years, 62.9% female). Abnormal functional testing had the highest specificity (94.2% (95% CI 88.4%-97.6%)) and positive likelihood ratio (4.55 (95% CI 1.86-11.13)) while CAC had the highest sensitivity (63.2% (95% CI 46.0%-78.2%)) and lowest negative likelihood ratio (0.52 (95% CI 0.34-0.80)). Specificity for ACC/AHA risk assessment was also high (88.5% (95% CI 81.1%-93.7%)). Over 3.3 years of follow-up, abnormal functional testing (adjusted subdistribution hazard ratio (SHR) 5.10, 95% CI 2.41 - 10.78, p < 0.001) and CAC (adjusted SHR 3.58, 95% CI 1.35 - 9.47, p = 0.010) were both significantly associated with MACE. CONCLUSIONS: In HL survivors treated with RT, both abnormal functional testing and ACC/AHA risk assessment had high specificity for subsequent MACE, but CAC had higher sensitivity. Further research is needed to inform CAD screening and primary prevention strategies in this population.

CRISPR-Cas9-mediated nuclear transport and genomic integration of nanostructured genes in human primary cells.

DNA nanostructures are a promising tool to deliver molecular payloads to cells. DNA origami structures, where long single-stranded DNA is folded into a compact nanostructure, present an attractive approach to package genes; however, effective delivery of genetic material into cell nuclei has remained a critical challenge. Here, we describe the use of DNA nanostructures encoding an intact human gene and a fluorescent protein encoding gene as compact templates for gene integration by CRISPR-mediated homology-directed repair (HDR). Our design includes CRISPR-Cas9 ribonucleoprotein binding sites on DNA nanostructures to increase shuttling into the nucleus. We demonstrate efficient shuttling and genomic integration of DNA nanostructures using transfection and electroporation. These nanostructured templates display lower toxicity and higher insertion efficiency compared to unstructured double-stranded DNA templates in human primary cells. Furthermore, our study validates virus-like particles as an efficient method of DNA nanostructure delivery, opening the possibility of delivering nanostructures in vivo to specific cell types. Together, these results provide new approaches to gene delivery with DNA nanostructures and establish their use as HDR templates, exploiting both their design features and their ability to encode genetic information. This work also opens a door to translate other DNA nanodevice functions, such as biosensing, into cell nuclei.

Role of Exercise Treadmill Testing in the Assessment of Coronary Microvascular Disease.

OBJECTIVES: The authors aimed to study the sensitivity and specificity of exercise treadmill testing (ETT) in the diagnosis of coronary microvascular disease (CMD), as well as the prognostic implications of ETT results in patients with CMD. BACKGROUND: ETT is validated to evaluate for flow-limiting coronary artery disease (CAD), however, little is known about its use for evaluating CMD. METHODS: We retrospectively studied 249 consecutive patients between 2006 and 2016 who underwent ETT and positron emission tomography within 12 months. Patients with obstructive CAD or left ventricular systolic dysfunction were excluded. CMD was defined as a coronary flow reserve <2. Patients were followed for the occurrence of a first major adverse event (composite of death or hospitalization for myocardial infarction or heart failure). RESULTS: The sensitivity and specificity of a positive ETT to detect CMD were 34.7% (95% CI: 25.4%-45.0%) and 64.9% (95% CI: 56.7%-72.5%), respectively. The specificity of a positive ETT to detect CMD increased to 86.8% (95% CI: 80.3%-91.7%) when only classifying studies with ischemic electrocardiogram changes that lasted at least 1 minute into recovery as positive, although at a cost of lower sensitivity (15.3%; 95% CI: 8.8%-24.0%). Over a median follow-up of 6.9 years (IQR: 5.1-8.2 years), 30 (12.1%) patients met the composite endpoint, including 13 (13.3%) with CMD (n = 98). In patients with CMD, ETT result was not associated with the composite endpoint (P = 0.076). CONCLUSIONS: Our data suggest limited sensitivity of ETT to detect CMD. However, a positive ETT with ischemic changes that persist at least 1 minute into recovery in the absence of obstructive CAD should raise suspicion for the presence of CMD given a high specificity. Further study is needed with larger patient sample sizes to assess the association between ETT results and outcomes in patients with CMD.

Clinical and Molecular Study of the NOG Gene in Families with Mandibular Micrognathism.

OBJECTIVES: Previous studies showed that noggin gene (NOG) sequence alterations, as well as epigenetic factors, could influence mandibular development. The aim of this study was to analyze clinical characteristics, NOG gene sequences, and promoter methylation sites in patients with mandibular micrognathism. MATERIALS AND METHODS: A total of 35 individuals of five Colombian families were subject to clinical and cephalometric analysis for mandibular micrognathism. One nonaffected individual of each family was included as a control. DNA was isolated from whole blood sample from all individuals by salting out method. Nine NOG gene fragments were amplified by polymerase chain reaction (PCR) and sequenced. Identification of CpG islands for methylation analysis at the NOG gene promoter was performed by MSP-PCR kit (Qiagen R). STATISTICAL ANALYSIS: A descriptive statistical analysis was carried out evaluating the presence or absence of genetics variants and the methylation sites in the NOG gene. RESULTS: NOG sequence results of affected individuals with mandibular micrognathism for one of the families studied demonstrated that they were heterozygous for 672 C/A (new mutation). For a second family, individuals were heterozygous for 567 G/C (single nucleotide polymorphism [SNP] RS116716909). For DNA analyzed from all patients studied, no methylations were observed at the NOG gene promoter region. CONCLUSION: Our results suggested that 672 C/A and 567 G/C variants could be involved in the presence of mandibular micrognathism. Moreover, lack of methylation sites at the NOG gene promoter region of all individuals studied suggests possibly other epigenetic factors could modulate mandibular growth. The search of genetic variants related with mandibular micrognathism will allow to predict in an integral way the development patterns of the patients and therefore establish a better clinical treatment.

Factors associated with adolescent HPV vaccination in the U.S.: A systematic review of reviews and multilevel framework to inform intervention development.

Multilevel factors impact HPV vaccine series initiation and completion among adolescents in the U.S. Synthesis of these factors is needed to inform intervention development and to direct future research. Current frameworks synthesizing factors focus on females only and do not include both series initiation and completion outcomes. We conducted a systematic review of reviews to identify modifiable individual-, provider-, and clinic-level factors associated with HPV vaccination outcomes among U.S. adolescents and developed a multilevel framework illustrating relations between factors to inform intervention development. We searched Medline, PsychInfo, Pubmed, CINAHL, and ERIC databases and included reviews published 2006 to July 2, 2018 describing individual-, provider-, or clinic-level factors quantitatively associated with HPV vaccination among U.S. adolescents. Two coders independently screened reviews, extracted data, and determined quality ratings. Sixteen reviews containing 481 unique primary studies met criteria. Factors synthesized into the multilevel framework included parent psychosocial factors (knowledge, beliefs, outcome expectations, intentions) and behaviors, provider recommendation, and patient-targeted and provider-targeted clinic systems. The scope of our framework and review advances research in two key ways. First, the framework illustrates salient modifiable factors at multiple levels on which to intervene to increase HPV vaccination. Second, the review identified critical gaps in the literature at each level. Future research should link the body of literature on parental intentions to vaccination outcomes, identify provider psychosocial factors associated with recommendation behaviors and subsequent vaccine uptake in their patient population, and understand clinic factors associated with successful implementation of patient- and provider-targeted system-level interventions.

Diagnóstico e intervención temprana de insuficiencia velofaringea posterior a palatoplastia primaria en pacientes atendidos interdisciplinariamente en fisulab (Bogotá, Colombia) / Diagnosis and early intervention of velofaringeal insufficiency posterior to primary palatoplasty in patients attended interdisciplinarily in fisulab (Bogotá, Colombia)

Introducción y Objetivo: El tratamiento integral e interdisciplinario de los pacientes con paladar hendido es fundamental para garantizar resultados a largo plazo. El habla es una de las metas principales en el tratamiento ya que permitirá un adecuado aprendizaje escolar y socialización. La insuficiencia velofaríngea (IVF) se debe a alteraciones anatómicas del esfínter velofaringeo y el manejo temprano y adecuado de los pacientes con paladar hendido es clave para evitarla. El objetivo de este estudio es determinar el porcentaje de IVF detectado y manejado antes de los primeros 5 años de vida en pacientes con paladar hendido tratados interdisciplinariamente en el Centro de Rehabilitación para Niños con Labio y Paladar Fisurado (FISULAB) en Bogotá, Colombia, evidenciando la efectividad del abordaje temprano. Material y Método: Estudio descriptivo tipo serie de casos en el que presentamos los resultados funcionales del tratamiento continuo en pacientes con paladar hendido, diagnosticados y tratados de forma exclusiva en FISULAB de acuerdo a su guía integral. Del total de 972 pacientes atendidos en la institución, 118 cumplían con los criterios de inclusión. Resultados: Estudiamos 118 pacientes con paladar hendido con o sin labio hendido atendidos bajo la guía integral desde el nacimiento hasta los 5 años de edad: 67 varones y 51 mujeres. Del total, 17 (14.4%) presentaron IVF (8 varones y 9 mujeres). De los que presentaron IVF, 4 (23.5%) tenían diagnóstico sindrómico asociado. La incidencia de IVF en nuestra población de pacientes, excluyendo los pacientes sindromáticos, fue del 11%, con una resolución del 100% de la IVF en los pacientes que fueron llevados a uno o múltiples procedimientos quirúrgicos. Conclusiones: El diagnóstico temprano y el manejo interdisciplinario de la IVF son esenciales para asegurar el éxito en la rehabilitación del lenguaje de los pacientes con paladar hendido. Un tratamiento conservador y por etapas de la IVF puede asegurar buenos resultados minimizando complicaciones

Background and Objective: A comprehensive and interdisciplinary treatment of patients with cleft palate is essential to ensure satisfactory long-term results. Adequate speech is one of the main goals because it allows each patient a proper adjustment to their educational and social environment. Velo- pharyngeal insufficiency (VPI) occurs due to anatomic changes of the velo- pharyngeal sphincter. Early and appropriate management of patients with cleft palate is crucial to prevent velopharyngeal insufficiency. The aim of this study is to determine the incidence of patients with VPI that were detected and received treatment during their first 5 years of age at FISULAB (Rehabilitation Center for Children with Cleft lip and Palate), Bogota, Colombia, and to evaluate the effectiveness early treatment. Methods: Descriptive study of case series. Patients included were diagnosed with cleft palate and treated exclusively in FISULAB, following the treatment protocol of this institution. A total of 972 patients were treated and 118 patients met the inclusion criteria. Results: A total of 118 patients with cleft palate, with or without associated cleft lip, were treated following the institutional protocol: 17 (14.4%) had VPI (8 male and 9 female). Of the patients who had VPI, 4 (23.5%) had a syndromic diagnosis. The incidence of VPI in our patient population was 11% with a resolution of 100% of VPI after surgical treatment. Conclusions: Early diagnosis and interdisciplinary management of VPI are essential to ensure the success in speech rehabilitation in patients with cleft palate. A conservative and stepwise treatment of VPI has shown good results, minimizing complications

El Tamaño del Plato no Afecta la Ingesta Energética de Personas con Sobrepeso y Obesidad / Plate size does not affect energy intake in obese or overweight persons

Se comparó el efecto de dos tamaños de plato sobre la ingesta energética en una situación de laboratorio. Los participantes fueron expuestos a platos de 26 cm y 22.5 cm de diámetro con 730 g de pasta disponible para comer lo que desearan en dos sesiones experimentales. En un segundo experimento fueron expuestos a las mismas condiciones con la excepción de que ellos debían servirse la cantidad que desearan comer. Se midió el IMC y la ingesta energética. Se aplicó la Escala Análoga Visual (EVA) para medir el nivel de saciedad. Los participantes con normopeso consumieron más alimento del plato pequeño que del plato grande, mientras que los participantes con sobrepeso y obesidad reportaron ingesta energética similar en todas las condiciones. No se encontró correlación entre el tamaño de platos y nivel de saciedad. Se discutieron los resultados con base en las estrategias para el control de la ingesta energética en personas con sobrepeso y obesidad

The effect of two dish sizes on a meal's energy intake in a laboratory situation was compared. Participants were exposed to a 26 cm/22.5 cm diameter dish with 730 g of pasta available to eat as they wished in two sessions. In the second experiment, participants were instructed to serve the food in one of the two sizes of a bowl in each session to eat what they wanted. Participants’ BMI was recorded and the energy intake was measured in each session. Additionally, the Visual Analogue Scale (VAS) was applied to measure participants' satiety level. Participants with normal weight consumed more food from the small plate than from the large plate, while participants with overweight and obesity consumed the same despite the difference in the size of plates. The results are discussed based on energy intake control strategies in over-weight and obese people

Using Intervention Mapping to Develop and Adapt Two Educational Interventions for Parents to Increase HPV Vaccination Among Hispanic Adolescents.

Introduction: Effective interventions to increase HPV vaccination are needed to reach national vaccination goals and to reduce later HPV-related cancer disparities. We used Intervention Mapping (IM) to develop and adapt a theory- and evidence-based educational intervention targeting parents of Hispanic adolescents to increase HPV vaccination. Methods: We followed IM steps 1-6 to: (1) develop a logic model and identify modifiable factors associated with vaccination among Hispanic adolescents by conducting literature reviews, focus groups, and in-depth interviews with Hispanic parents; (2) develop outcomes, write performance objectives, and develop a matrix of change objectives; (3) develop and identify a program theme, program components, theoretical methods, and practical applications; (4) develop an intervention design plan; (5) develop implementation strategies; and (6) develop an evaluation plan. We completed Steps 1-6 for to develop an intervention targeting parents of females, and we followed the steps again to adapt the program once HPV vaccine recommendations included males. Results: The program Por Nuestras Hijas (For Our Daughters) included two components: a print fotonovela and a tailored interactive multimedia intervention (TIMI). The program utilized the methods tailoring, targeting, framing, anticipated regret, modeling, skill building, and education and counseling to target the following determinants: parental knowledge, attitudes, self-efficacy, skills, perceived benefits/barriers, perceived susceptibility, perceived norms, and outcome expectations as modifiable factors influencing HPV vaccination. Lay health workers implemented the program in community clinics. A logic model of change guided evaluation planning. We later adapted the outcome and intervention content for parents of Hispanic adolescent males and changed the theme to Por Nuestros Hijos (For Our Children). Throughout the development and adaptation processes, we relied on theory, empirical evidence, and new data to make decisions. Discussion: IM provided a systematic methodology for program development and adaptation. Tasks in each step built upon one another integrating findings from the literature, previous research, qualitative findings, and theory to develop two educational programs for parents to increase HPV vaccination. The systematic process allowed us to develop messages and materials targeting factors beyond HPV knowledge or awareness to create behavior change.

Influencia de la luz azul sobre la productividad del cultivo sólido de Ganoderma lucidum / Influence of blue light on the productivity of the solid culture of Ganoderma lucidum

RESUMEN Ganoderma lucidum es un hongo macromiceto reconocido por sus propiedades medicinales y su contenido de compuestos bioactivos que incluyen polisacáridos, triterpenoides, proteínas inmunomoduladoras, entre otros, lo que ha generado un incremento notable en su producción. La mayoría de especies de hongos responden y se adaptan a diversas señales ambientales incluida la luz, que favorece su productividad, tanto en calidad como en cantidad al estar estrechamente relacionada con la formación de cuerpos fructíferos. Por tal razón, el objetivo de este estudio fue evaluar la eficiencia biológica (EB) y la tasa de producción (TP) como parámetros de productividad del cultivo sólido de Ganoderma lucidum bajo irradiación de los sustratos con luz emitida por diodos azules (LED) con dos periodos de foto-estímulo de 12 y 24 h durante todas las fases de cultivo para inducir el crecimiento micelial y la formación de los cuerpos fructíferos. Se aplicaron parámetros convencionales para el crecimiento y desarrollo del hongo en las etapas de producción. Para la formulación de los sustratos, se emplearon residuos agroindustriales y materiales lignocelulósicos. El diámetro de los cuerpos fructíferos sometidos a tratamientos con luz azul fue mayor que los exhibidos a luz blanca fluorescente (Testigo). Los resultados muestran que el cultivo de Ganoderma lucidum con exposición a la luz azul es útil para la inducción de cuerpos fructíferos de alta calidad, logrando una disminución del periodo de fermentación en 16 días para el foto-estímulo de 24 h con EB de 28,04% y TP de 0,64.

ABSTRACT Ganoderma lucidum, a renowned fungus for its medicinal properties and content of bioactive compounds include polysaccharides, triterpenoids, immune modulatory proteins, among others; it has generated a significant increase and interest in its production. Most species of fungi respond and adapt to various environmental signal including light, which affects it not only in productivity, but also in quality and quantity to be closely related to the fruiting bodies. Therefore, the objective of this study was evaluating the biological efficiency (EB) and the rate of production (TP) as production parameters of the solid culture of Ganoderma lucidum at under irradiation of the substrate with light emitting diodes blue (LED) with two levels of photo-stimulation constant 12 and 24 h during all steps of culture to induce mycelial growth and the formation of fruiting bodies. Conventional parameters were applied to stimulate the growth and development of the fungus in the stages of production. For the formulation of substrates agro-industrial waste and lignocellulosic materials were used. The diameter of the fruiting bodies under blue light treatments was higher than those exposed to white fluorescent light (control). The results show that the cultivation of Ganoderma lucidum with exposure to blue light is useful for induction of fruiting bodies of high quality, achieving a reduction of the period of fermentation in 16 days for the photo-stimulation of 24 h with EB of 28,04% and TP of 0,64.

Initiation of inflammatory tumorigenesis by CTLA4 insufficiency due to type 2 cytokines.

Genetically predisposed CTLA4 insufficiency in humans is associated with gastric cancer development, which is paradoxical to the prototypical role of CTLA4 in suppressing antitumor immunity. CTLA4 is a critical immune checkpoint against autoimmune disorders. Autoimmunity has been implicated in protumor or antitumor activities. Here, we show that CTLA4 insufficiency initiates de novo tumorigenesis in the mouse stomach through inflammation triggered by host-intrinsic immune dysregulation rather than microbiota, with age-associated progression to malignancy accompanied by epigenetic dysregulation. The inflammatory tumorigenesis required CD4 T cells, but not the TH1 or TH17 subsets. Deficiencies in IL-4 and IL-13 or IL-4 receptor α broke the link between inflammation and initiation of tumorigenesis. This study establishes the causality of CTLA4 insufficiency in gastric cancer and uncovers a role of type 2 inflammation in initiating gastric epithelial transformation. These findings suggest possible improvement of immune therapies by blocking tumorigenic type 2 inflammation while preserving antitumor type 1 immunity.

Ammonia and greenhouse gases losses from mechanically turned cattle manure windrows: A regional composting network.

An on-farm composting network operates in the Basque Country (northern Spain), in which solid manure produced in livestock farms (mostly dairy and beef cattle) is composted through windrow turning. This network aims to produce a valuable resource (compost) for the farmers whereas the volume of the solid manure was reduced at farm level The objective of the study was to assess the gaseous losses (NH3 and GHG) from 6 on-farm composting windrows (either deep litter systems or solid fraction after slurry separation) after turning operations. Monitored turning events occurred 1 to 4 months after establishing the heaps on the field. Ammonia and greenhouse gas (GHG) losses were estimated by the open and close chamber techniques, respectively. Results showed overall low emission rates related to the long degradation period of the windrows. Maximum NH3 release was at 2.0 mg m-2 d-1 after the second/third turning events. Baseline N2O losses were below 50 mg m-2 d-1, with maximum rates close to 500 mg m-2 d-1 some days after turning works. Methane emissions were mostly below 100 mg m-2 d-1, while CO2 losses were lower than 25 g m-2 d-1. Carbon dioxide peaks (≈250 g m-2 d-1) were reached after the second/third turnings. Overall, gaseous N and C losses accounted for 0.1 and 1% of the initial N and C content of the windrows, respectively. The present study concluded that two/three turning operations in aged solid manure-derived compost windrows do not have significant effects on NH3 and GHG losses. The magnitude of the gaseous losses from on-farm composting systems is dependent on the manure management practices at farm level (e.g. moment of windrow stacking).

Orbital Reconstruction in Neuroectodermal Tumor of the Orbit: Multimodal Treatment Approach.

INTRODUCTION: Primitive neuroectodermal tumors of peripheral origin are very rare, and orbital neuroectodermal tumors are even more uncommon. Only 25 patients with primary orbital involvement in the pediatric age group have been reported. METHODS: In this article, the authors describe their experience in the multimodality treatment approach to treat neuroectodermal tumor of the orbit. The authors also present a male patient 3-year old presenting with a neuroectodermal tumor of the right orbit causing rapidly progressive proptosis. The patient underwent an upper and lateral orbital marginotomy. The entire bone defect was reconstructed with a bone graft, allowing for the reconstruction of the floor and the lateral wall of the middle cranial fossa, the floor of the anterior cranial fossa, the upper and lateral orbital frame, and the right zygomatic bone. Over a period of 16 months, the patient was subjected to chemotherapy. RESULTS: In the postoperative period, a favorable evolution of the disease was observed, with growth in the reconstructed structures, good projection of the orbit and the eyeball, and stable results without tumor recurrence. CONCLUSIONS: The authors present the clinical analysis, surgical management, as well as the chemotherapy treatment established, with follow-ups at 1 and 2 and a half years. This experience shows the effectiveness of multimodality therapy in the treatment of rare tumors of difficult handling.

Adipocyte and leptin accumulation in tumor-induced thymic involution.

Cell-mediated immunity is an important defense mechanism against pathogens and developing tumor cells. The thymus is the main lymphoid organ involved in the formation of the cell-mediated immune response by the maturation and differentiation of lymphocytes that travel from the bone marrow, through the lymphatic ducts, to become T lymphocytes. Thymic involution has been associated with aging; however, other factors such as obesity, viral infection and tumor development have been shown to increase the rate of shrinkage of this organ. The heavy infiltration of adipocyte fat cells has been reported in the involuted thymuses of aged mice. In the present study, the possible accumulation of such cells in the thymus during tumorigenesis was examined by immunohistochemistry. A significant number of adipocytes around and infiltrating the thymuses of tumor-bearing mice was observed. Leptin is a pro-inflammatory adipocytokine that enhances thymopoiesis and modulates T cell immune responses. The levels of leptin and adiponectin, another adipocytokine that has anti-inflammatory properties, were examined by western blot analysis. While no changes were observed in the amounts of adiponectin present in the thymuses of the normal and tumor-bearing mice, significantly higher levels of leptin were detected in the thymocytes of the tumor-bearing mice. This correlated with an increase in the expression of certain cytokines, such as interleukin (IL)-2, interferon (IFN)-γ and granulocyte-macrophage colony-stimulating factor (GM-CSF). The co-culture of thymocytes isolated from normal mice with ex vivo isolated adipocytes from tumor-bearing mice yielded similar results. Our findings suggest that the infiltration and accumulation of adipocytes in the thymuses of tumor-bearing mice play an important role in their altered morphology and functions.

Insights into thymic involution in tumor-bearing mice.

The thymus is a central lymphoid organ critical for the development and maintenance of an effective peripheral T-cell repertoire. Most important, it provides a specialized environment for the selection of rearranged clones that will function appropriately in the adaptive immune response. Thymic involution has been observed in several model systems; including graft-versus-host disease, aging, viral infection, and tumor development, however, the precise mechanisms involved in this phenomenon remain poorly defined. Here, we review some of our results related to the studies of the cell-mediated immunity in a mammary tumor model; more specifically, those related to the tumor-induced impaired T-cell development and thymic involution. Collectively, the understanding of the mechanisms and pathways associated with the tumor-induced thymic involution is essential for the development of innovative and safe therapies to fight against the immune suppression caused by the tumor development.

Mechanisms of antitumor and immune-enhancing activities of MUC1/sec, a secreted form of mucin-1.

Mucin 1 (MUC1) is a polymorphic type 1 transmembrane protein found on the apical surface of normal cells lining the lumen of ducts and glands. Mucins are thought to provide mucosal protection from environmental exposures and carcinogens. An altered form of the MUC1 glycoprotein, which is hypoglycosylated, is expressed in several types of human cancers. In our laboratory, we have found that transfection of a murine mammary tumor cell line with a human secreted isoform of MUC1 rendered these DA-3 cells (DA-3/sec) incapable of growing in intact BALB/c mice. In contrast, implantation of DA-3 cells transfected with the human transmembrane isoform of MUC1 (DA-3/TM), resulted in tumor formation and ultimately death of the animals, similar to the DA-3 parental line. Importantly, inoculation of the DA-3/sec cells in immunodeficient nude mice resulted in tumor formation, indicating that the MUC1/sec molecule's antitumor activity is immunologically controlled. In this review, we summarize the studies we have performed to elucidate possible mechanisms for the immune-mediated antitumor effect of MUC1/sec and/or a unique peptide present in this mucin. Understanding these mechanisms may provide new immunotherapeutic approaches that could be used to target different types of cancer.

uPA and uPA-receptor are involved in cancer-associated myeloid-derived suppressor cell accumulation.

BACKGROUND: Myeloid-derived suppressor cells (MDSC) have been shown to play a critical role in tumor-induced immunosuppression, in many mouse and human cancers. The aim of this study was to show that MDSC accumulation is tumor burden-dependent, and to investigate the role of the tumor-derived urokinase plasminogen activator (uPA) and its receptor (uPAR) on MDSC recruitment. MATERIALS AND METHODS: Levels of MDSC were assessed in tumor-bearers, and the ability to recruit MDSC by uPA was investigated in normal, tumor-bearers, uPAR(-/-), and CD11b(-/-) mice. uPAR expression in MDSC was also explored. RESULTS: MDSC accumulate to dramatic levels in tumor-bearers, and tumor-derived factors such as uPA also increase to great levels in circulation. MDSC can be recruited by uPA, and uPAR but not CD11b are required for such recruitment. CONCLUSION: MDSC accumulation is tumor burden-dependent, and tumor-derived factors such as uPA and its receptor uPAR play a role in their recruitment.

Decreased accumulation of immune regulatory cells is correlated to the antitumor effect of IFN-γ overexpression in the tumor.

During mammary tumorigenesis, there is a profound tumor-induced immunosuppression and a progressive thymic atrophy associated with tumor development. IFN-γ has been shown to be effective in enhancing antitumor responses in several tumor models, however, how IFN-γ exerts its anti-tumor effect is largely controversial. In the present study we have used a mammary tumor model to investigate whether the levels of IFN-γ have an important role in the tumor-induced immuno-suppression as well as in the pathogenesis of the thymic atrophy. We evaluated this possibility using DA-3 cells transfected to express IFN-γ (DA-3/IFN-γ), a system that provides constant, local production of IFN-γ within the tumor microenvironment. Overexpression of IFN-γ in the mammary tumor results in a marked delay of tumor growth, a reduction in regulatory T cells and myeloid-derived suppressor cells accumulation mostly due to down-regulation of chemokines implicated in the recruitment of immune regulatory cells, and a blockage in the tumor-associated thymus atrophy. Collectively, our data suggest that the replacement of the faulty levels of IFN-γ in the tumor results in a diminution of the tumor-induced immune suppression caused by the mammary tumor development.

Tumor-induced thymic atrophy: alteration in interferons and Jak/Stats signaling pathways.

The thymus is the major site of T cell differentiation and a key organ of the immune system. Thym atrophy has been observed in several model systems including aging, and tumor development. Previous results from our laboratory have reported that the thymic atrophy seen in mammary tumor bearers is associated with a severe depletion of CD4+CD8+ double positive immature cells and changes in the levels of cytokines expressed in the thymus microenvironment. Cytokines regulate numerous aspects of hematopoiesis via activation of the Jak/Stat pathways. In the present study we have used our mammary tumor model to investigate whether changes in the levels of cytokines in the thymus could affect the normal expression of the aforementioned pathways. RNA and protein analysis revealed an overexpression of the different members of interferons, a downregulation of most of the Jak/Stat pathways, and an increased expression of several suppressors of cytokine signaling (SOSC) in the thymuses of tumor bearers. Together, our data suggest that the impaired Jak/Stat signaling pathways observed in the whole thymus of tumor-bearing mice could be contributing to the abnormal T cell development and apoptosis observed during the tumor-induced thymic atrophy.

The liver is a site for tumor-induced myeloid-derived suppressor cell accumulation and immunosuppression.

Tumor-induced immunosuppression plays a key role in tumor evasion of the immune system. A key cell population recognized as myeloid-derived suppressor cells (MDSC) contributes and helps orchestrate this immunosuppression. MDSC can interact with T cells, macrophages, and natural killer cells to create an environment favorable for tumor progression. In various tumor models, their presence at high levels has been reported in the bone marrow, blood, spleen, and tumor. We report for the first time that MDSC accumulate and home to the liver in addition to the other organs. Liver MDSC suppress T cells and accumulate to levels comparable with splenic MDSC. Additionally, hematopoiesis in the liver contributes to the dramatic expansion of MDSC in this organ. Furthermore, MDSC in the liver interact with macrophages, also known as Kupffer cells, and cause their up-regulation of PD-L1, a negative T-cell costimulatory molecule. The liver is thus an organ in which MDSC accumulate and can contribute to immunosuppression directly and indirectly. MDSC play a role in various pathologic states in addition to cancer, and these results contribute to our understanding of their biology and interactions with immune-related cells.

Decreased levels of both Stat1 and Stat3 in T lymphocytes from mice bearing mammary tumors.

BACKGROUND: Lymphocytes from tumor-bearing animals have been shown to lack antitumor function. The objective of this study was to investigate the status of the signal transducers, Stat1 and Stat3, in T lymphocytes of animals bearing D1-DMBA-3 mammary tumors and to elucidate if any alterations in these signal transducers can be explained by the presence of tumor-derived factors and correlated with the lack of antitumor function in these cells. MATERIALS AND METHODS: T Lymphocytes from spleens of normal and tumor-bearing mice were purified and assayed for the presence of Stat1 and Stat3 by Western blot analysis. RESULTS: It was found that levels of both Stat1 and Stat3 were reduced in T lymphocytes of tumor-bearers not only in their active, phosphorylated form but in total protein levels. CONCLUSION: These findings indicate that during mammary tumor progression, alteration of various transcription factors may contribute to the down-regulation of immune function.