Search

Supplementary Figure from NR2F1 Is a Barrier to Dissemination of Early-Stage Breast Cancer Cells

Cancer cells can disseminate during very early and sometimes asymptomatic stages of tumor progression. Though biological barriers to tumorigenesis have been identified and characterized, the mechanisms that limit early dissemination remain largely unknown. We report here that the orphan nuclear receptor nuclear receptor subfamily 2, group F, member 1 (NR2F1)/COUP-TF1 serves as a barrier to early dissemination. NR2F1 expression was decreased in patient ductal carcinoma in situ (DCIS) samples. High-resolution intravital imaging of HER2+ early-stage cancer cells revealed that loss of function of NR2F1 increased in vivo dissemination and was accompanied by decreased E-cadherin expression, activation of wingless-type MMTV integration site family, member 1 (WNT)-dependent β-catenin signaling, disorganized laminin 5 deposition, and increased expression of epithelial–mesenchymal transition (EMT) genes such as twist basic helix-loop-helix transcription factor 1 (TWIST1), zinc finger E-box binding homeobox 1 (ZEB1), and paired related homeobox 1 (PRRX1). Furthermore, downregulation of NR2F1 promoted a hybrid luminal/basal phenotype. NR2F1 expression was positively regulated by p38α signaling and repressed by HER2 and WNT4 pathways. Finally, early cancer cells with NR2F1LOW/PRRX1HIGH staining were observed in DCIS samples. Together, these findings reveal the existence of an inhibitory mechanism of dissemination regulated by NR2F1 in early-stage breast cancer cells.Significance:During early stages of breast cancer progression, HER2-mediated suppression of NR2F1 promotes dissemination by inducing EMT and a hybrid luminal/basal-like program.

Supplementary Data from NR2F1 Is a Barrier to Dissemination of Early-Stage Breast Cancer Cells

Cancer cells can disseminate during very early and sometimes asymptomatic stages of tumor progression. Though biological barriers to tumorigenesis have been identified and characterized, the mechanisms that limit early dissemination remain largely unknown. We report here that the orphan nuclear receptor nuclear receptor subfamily 2, group F, member 1 (NR2F1)/COUP-TF1 serves as a barrier to early dissemination. NR2F1 expression was decreased in patient ductal carcinoma in situ (DCIS) samples. High-resolution intravital imaging of HER2+ early-stage cancer cells revealed that loss of function of NR2F1 increased in vivo dissemination and was accompanied by decreased E-cadherin expression, activation of wingless-type MMTV integration site family, member 1 (WNT)-dependent β-catenin signaling, disorganized laminin 5 deposition, and increased expression of epithelial–mesenchymal transition (EMT) genes such as twist basic helix-loop-helix transcription factor 1 (TWIST1), zinc finger E-box binding homeobox 1 (ZEB1), and paired related homeobox 1 (PRRX1). Furthermore, downregulation of NR2F1 promoted a hybrid luminal/basal phenotype. NR2F1 expression was positively regulated by p38α signaling and repressed by HER2 and WNT4 pathways. Finally, early cancer cells with NR2F1LOW/PRRX1HIGH staining were observed in DCIS samples. Together, these findings reveal the existence of an inhibitory mechanism of dissemination regulated by NR2F1 in early-stage breast cancer cells. Significance: During early stages of breast cancer progression, HER2-mediated suppression of NR2F1 promotes dissemination by inducing EMT and a hybrid luminal/basal-like program.

Pre-malignant cells disseminate during very early and sometimes asymptomatic stages of tumor progression. Once settled in permissive environments at secondary organs, they can act as a source for metastasis (e.g. in breast, pancreatic, colon, melanoma and lung carcinomas), thus emerging as important players in future prognosis. Granted that biological barriers to tumorigenesis exist, there must also be limiting steps to early dissemination in pre-malignant cells, all of which remain largely unknown. We report that the orphan nuclear receptor NR2F1/COUP-TF1, downstream of Her2 signaling via p38alpha serves as a barrier to early dissemination in pre-malignant breast cells. Our work shows that NR2F1 expression is positively regulated by p38α-MAPK pathway and inhibited by HER2 oncogenic signaling pathway. Furthermore, NR2F1 downregulation in human and murine pre-malignant cell lines led to a partial epithelial-to-mesenchymal transition characterized by decreased E-cadherin expression and β-catenin membrane localization, disorganized laminin 5 deposition, and increased expression of CK14, TWIST1, ZEB1 and PRRX1, thus explaining an increased motility and dissemination of early cells upon loss of NR2F1. Interestingly, NR2F1 lost also maintained epithelial markers (like CK18), suggesting an hybrid luminal-mesenchymal phenotype that could favor cellular plasticity to adapt to distant sites. We also showed that NR2F1 blocked b-catenin nuclear localization via a WNT-dependent pathway. Significantly, downregulation of NR2F1 increased the in vivo dissemination potential of pre-malignant cells to lungs in a HER2-dependent manner. Lastly, Nr2f1 expression was downregulated in non-invasive DCIS biopsies when compared to benign adjacent tissue, and its expression was inversely correlated to Prrx1 levels. The inverse correlation between Nr2f1 and Prrx1 levels was lost in genetically evolved primary tumors, which suggests that a unique inhibitory mechanism of dissemination dominates at early stages of tumor progression. Overall, our data suggest the existence of an inhibitory mechanism of dissemination regulated by NR2F1 downstream of HER2 signaling in premalignant breast cells and propose that therapeutic strategies that aim to maintain NR2F1 expression could reduce dissemination of premalignant cells and delay relapses. Citation Format: Carolina Rodriguez-Tirado, Nupura Kale, Maria Jose Carlini, Nitisha Shrivastava, Alcina Rodrigues, Bassem Khalil, Jose Javier Bravo-Cordero, Melissa Alexander, Jiayi Ji, Maria Soledad Sosa. NR2F1 is a barrier to dissemination of early-evolved mammary cancer cells [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P3-06-03.

Objective: To analyze and find risk factors associated with the development of DI using a multicenter case series via TSE. Materials and methods: Medical records of patients who underwent TSS for PA resection between 2010-2021 at three different neurosurgical centers by four experienced neurosurgeons were retrospectively analyzed. The patients were divided into two groups (DI Group or Control Group). Logistic regression analysis was conducted to identify risk factors associated with postoperative DI. Univariate logistic regression was performed to identify variables of interest, and covariates with a p-value < 0.15 were incorporated into multivariate logistic regression models to identify independently associated risk factors for DI. All statistical tests were conducted using RStudio. Results: 344 patients were included; 68% were women, the mean age was 46.5 years, and non-functioning adenomas were the most frequent (171, 49.7%). The mean tumor size was 20.3 mm. Covariates associated with postoperative DI were female gender, cavernous sinus invasion (left or bilateral), intraoperative CSF leak, endoscopic surgery, left preoperative visual field involvement, complete tumor resection, Knosp score of 2 and 4, and macroadenomas. Age was associated with a lower OR for postoperative DI. Multivariable model showed female gender (OR 3.34 CI 1.50-7.99, p=0.004) and intraoperative CSF leak (OR 3.25 CI 1.25-8.85, p=0.017) were found to be independently associated with higher OR of developing postoperative DI. Age was not shown to be a protective factor for DI (OR 0.97; CI: 0.94-1.00; p=0.038). Conclusions: The independent risk factors for the development of DI were female patients and intraoperative CSF leak.

Since their approval in 2011, immune checkpoint inhibitors (ICPis) are increasingly used to treat several advanced cancers. ICPis target certain cellular molecules that regulate immune response resulting in antitumor activity. The use of these new agents needs careful monitoring since they brought a whole new spectrum of adverse events. In this review, we aim to describe different endocrine dysfunctions induced by ICPis and to underline the importance of diagnosing and managing these adverse effects. Immune-related endocrine toxicities include thyroid dysfunction, hypophysitis and, less frequently, type 1 diabetes, primary adrenal insufficiency and hypoparathyroidism. Diagnosis of endocrine adverse events related to ICPis therapy can be challenging due to nonspecific manifestations in an oncological scenario and difficulties in the biochemical evaluation. Despite the fact that these endocrine adverse events could lead to life-threatening consequences, the availability of effective replacement treatment enables continuing therapy and together with an interdisciplinary approach will impact positively on survival.

After a stressful event, adaptative mechanisms are carried out to support vital functions. Hypothalamic-pituitary-adrenal axis plays a key role in stress response regulating metabolism, cardiovascular function and immune system. This review addresses pathophysiological changes of the adrenal axis during critical illness, recognizing limitations of methods applied for its evaluation in this special context and defining indications for corticosteroid replacement in critically ill patients. The concept of relative adrenal insufficiency should be abandoned; cosyntropin stimulation test should not be performed for diagnosis of adrenal insufficiency in critical illness nor for establishing the need of treatment.Diversos mecanismos adaptativos se ponen en marcha para sostener las funciones orgánicas vitales en el paciente crítico. El eje hipotálamo-hipófiso-adrenal tiene un papel clave en la respuesta al estrés al regular el metabolismo, la función cardiovascular y la respuesta inmune. Esta revisión tiene por objetivos analizar los cambios fisiopatológicos que se producen en el eje adrenal durante la enfermedad crítica, reconocer las limitaciones de los métodos diagnósticos y definir indicaciones de tratamiento de reemplazo corticoideo en este contexto. El concepto de insuficiencia adrenal relativa debe ser descartado y no se recomienda el test de estímulo con cosintropina para diagnóstico de insuficiencia adrenal durante enfermedad crítica ni para definir la necesidad de tratamiento.

Conocer el impacto del aire producido por fuentes móviles, sigue siendo un desafío importante. En la mayoría de las ciudades argentinas, se desconoce el estado actual de la calidad del aire. Investigaciones dirigidas a conocer el aporte de contaminantes a partir de fuentes móviles, se han realizado para grandes ciudades, por ejemplo: Ciudad Autónoma de Buenos Aires, Mendoza, o Rosario. La complejidad del trabajo con diferentes flotas vehiculares (diversidad de tecnologías) y la dificultad en el acceso a la información en suficiente cantidad y calidad, son barreras para el avance de este tipo de estudios diagnósticos en ciudades intermedias o chicas. Este estudio tiene como objetivo principal inventariar y diagnosticar las emisiones de gases de efecto invernadero (GEI), y otros contaminantes (tóxicos y criterio) correspondientes a la categoría vehicular taxi/remis en la ciudad de Tandil, Buenos aires -Argentina, para el año 2016. Como herramienta metodológica se aplica el Modelo Internacional de Emisiones Vehiculares (IVE). El estudio caracteriza la flota y hace estimaciones de emisiones de los contaminantes que representan un problema para la salud por lo tanto afectan a la calidad del aire y pueden tener consecuencias para el cambio climático. En Tandil los taxis/remís generan al año un total de 9.231,99 Tn de CO2eq, mientras que los contaminantes criterio y tóxicos no representan un riesgo para la salud o el ambiente urbano debido al uso de GNC como principal combustible. El inventario es una herramienta fundamental en la gestión de la calidad del aire, este estudio presenta una línea de base para diagnosticar el estado actual de las emisiones y poder reflexionar sobre la tendencia de las mismas a partir del seguimiento en nuevos estudios. Abstract Know the impact of air produced by mobile sources continues to be a big challenge. In most Argentine cities, the current state of air quality is unknown. Investigations aimed at knowing the contribution of pollutants from mobile sources have been carried out for large cities, for example: City of Buenos Aires, Mendoza, or Rosario. The complexity of the work with different vehicle fleets (diversity of technologies) and the difficulty in accessing information in sufficient quantity and quality are barriers to the advancement of this type of diagnostic studies in intermediate or small cities. The main objective of this study is to inventory and diagnose GHG emissions and other pollutants (toxic and criteria) corresponding to the vehicle category taxi / remis in the city of Tandil, Buenos Aires -Argentina, for the year 2016. As a methodological tool, applies the International Model of Vehicle Emissions (IVE). The study characterizes the fleet and makes estimates of emissions of pollutants that represent a health problem and therefore affect air quality and may have consequences for climate change. In Tandil, taxis / remis generate a total of 9,231.99 Tn of CO2eq per year, while criteria and toxic pollutants do not represent a risk to health or the urban environment due to the use of CNG as the main fuel. The inventory is a fundamental tool in the management of air quality, this study presents a baseline to diagnose the current state of emissions and to be able to reflect on their trend from the follow-up in new studies.

Background Acromegaly is associated with higher morbidity and mortality mainly due to cardiovascular disease. Data on the incidence and evolution of thyroid cancer in acromegaly are controversial. Our objective was to describe the characteristics of a group of acromegalic patients with differentiated thyroid carcinoma (DTC) and analyze their evolution. Methods This is a retrospective multicenter study of 24 acromegalic patients with DTC. The AJCC Staging System 8th Edition was used for TNM staging, and the initial risk of recurrence (RR), initial response and response at the end of follow-up (RFU) were defined according to the 2015 ATA Guidelines. As a control group, 92 patients with DTC without acromegaly were randomly included. Statistical analyses were done using SPSS Statistics 20.0. Results Median age of patients at diagnosis of acromegaly was 49.5 years (range 12–69). The median delay in diagnosis of acromegaly was 3 years (range 0.5–23). Mean baseline IGF-1 level was 2.9 ± 1.1 ULN. Median age at DTC diagnosis was 51.5 years (18–69). At the moment of diagnosis of DTC, 58.3% of the patients had active acromegaly. Median time from DTC diagnosis to acromegaly control was 1.25 years (0.5–7). Mean DTC tumor diameter of the biggest lesion was 14.6 ± 9.2 mm, being multifocal in 37.5%. All tumors were papillary carcinomas, two cases being of an aggressive variety. Lymph node dissection was performed in 8 out of 24 patients and 62.5% had metastases. Only one patient had distant metastases. Radioiodine ablation was given to 87.5% of patients. Nineteen patients (79%) were stage I, four (17%) stage II and one (4%) stage IVb. Initial RR was low in 87% (21/24), intermediate in 9% (2/24) and high in 4% (1/24) patient. RFU was: 83% (19/23) patients with no evidence of disease, 9% (2/23) with indeterminate response, 4% (1/23) with biochemical incomplete response and 4% (1/23) with structural incomplete response, at a median time of FU of 36.5 months. When comparing RFU between acromegalics and controls no statistically significant differences were found. Conclusions Patients with acromegaly and DTC mostly had a low initial RR. When compared with the control group, we found that DTC patients with acromegaly did not have a worse evolution.

Lithium carbonate is a commonly prescribed drug for bipolar disorders. In addition to its action on the central nervous system, lithium has systemic effects on multiple organs such as kidney, heart, motor end plate, thyroid and parathyroid glands. It can cause hypothyroidism, hyperthyroidism, goiter and ophthalmopathy by different mechanisms. It increases intrathyroid iodine content or compete for iodine transport, resulting in low iodine uptake by the thyroid. It also inhibits the coupling of iodotyrosine residues to form iodothyronines and inhibits the release of T4 and T3. Lithium has direct actions on parathyroid glands by antagonizing the calcium sensing receptor, which may induce hypercalcemia or even hyperparathyroidism, requiring surgery in some cases. Furthermore, it inhibits the expression of aquaporins, mainly aquaporin 2, in the renal collecting tubule by unknown mechanisms leading to nephrogenic diabetes insipidus. This adverse effect is usually reversible after drug withdrawal. However, some patients may present irreversible kidney damage due to chronic interstitial nephropathy.El carbonato de litio es un fármaco que se prescribe comúnmente para el tratamiento de trastornos bipolares. Además de su acción sobre el sistema nervioso central, el litio tiene repercusiones sistémicas, afectando a múltiples órganos como el riñón, el corazón, la placa motora terminal y glándulas tiroides y paratiroides. Puede causar hipotiroidismo, hipertiroidismo, bocio y oftalmopatía por diferentes mecanismos; también aumentar el contenido de yodo intratiroideo o competir por el transporte de yodo, lo que resulta en una baja captación tiroidea de yodo. Inhibe el acoplamiento de residuos de yodotirosina para formar yodotironinas e inhibe la liberación de T4 y T3. Tiene acciones directas sobre las glándulas paratiroides antagonizando el receptor sensor de calcio, lo que puede inducir hipercalcemia e incluso hiperparatiroidismo, y puede requerir cirugía en algunos casos. Inhibe la expresión de acuaporinas en el túbulo colector renal, principalmente acuaporina 2, por mecanismos que aún no se conocen, produciendo diabetes insípida nefrogénica; este efecto adverso suele ser reversible tras la suspensión del fármaco. Sin embargo, algunos pacientes pueden presentar daño renal irreversible por nefropatía intersticial crónica.

Disseminated cancer cells (DCCs) identified in secondary organs, sometimes before the primary tumor becomes detectable and treated, can remain dormant for years to decades before manifesting. Microenvironmental and epigenetic mechanisms may control the onset and escape from dormancy, and here we reveal how a combination of the DNA methylation inhibitor 5-azacytidine (AZA) and retinoic acid receptor ligands all-trans retinoic acid (atRA), orchestrate a novel program of stable dormancy. Treatment of HNSCC tumor cells with AZA+atRA induced a SMAD2/3/4 dependent regulation of downstream transcriptional program that restored the anti-proliferative function of TGFβ signaling. Significantly, AZA+atRA or AZA+AM80, an RARα specific agonist, strongly suppresses lung metastasis formation. The metastatic suppression occurs via the induction and maintenance of phenotypically homogenous dormant SMAD4+/NR2F1+ non-proliferative DCCs. These findings suggest that strategies that maintain or induce dormancy programs may be a viable alternative strategy to improve patient outcomes by preventing or significantly delaying metastasis development.

We describe the discovery of an agonist of the nuclear receptor NR2F1 that specifically activates dormancy programs in malignant cells. The agonist led to a self-regulated increase in NR2F1 mRNA and protein and downstream transcription of a novel dormancy program. This program led to growth arrest of an HNSCC PDX line, human cell lines, and patient-derived organoids in 3D cultures and in vivo. This effect was lost when NR2F1 was knocked out by CRISPR-Cas9. RNA sequencing revealed that agonist treatment induces transcriptional changes associated with inhibition of cell cycle progression and mTOR signaling, metastasis suppression, and induction of a neural crest lineage program. In mice, agonist treatment resulted in inhibition of lung HNSCC metastasis, even after cessation of the treatment, where disseminated tumor cells displayed an NR2F1hi/p27hi/Ki-67lo/p-S6lo phenotype and remained in a dormant single-cell state. Our work provides proof of principle supporting the use of NR2F1 agonists to induce dormancy as a therapeutic strategy to prevent metastasis.

Disseminated tumor cells (DTCs) in secondary organs often remain dormant for a long period of time before re-awakening and growing into overt metastases. We have previously identified NR2F1/COUP-TF1, an orphan nuclear receptor, as a master regulator of tumor cell dormancy in head and neck squamous cell carcinoma (HNSCC) and other cancer types. Here we describe the identification and function of a novel NR2F1 agonist herein referred to as compound 26 (C26). C26 was found to specifically activate NR2F1 in HNSCC cells, leading to increased NR2F1 transcription and nuclear protein accumulation. C26-mediated activation of NR2F1 induced growth arrest of HNSCC PDX line and cell lines in 3D cultures in vitro and on chicken embryo chorioallantoic membrane (CAM) in vivo. The effect of C26 on growth arrest was lost when NR2F1 was knocked out by CRISPR/Cas9. C26-induced growth arrest was mediated by activation of an NR2F1-regulated dormancy program, including upregulation of cyclin-dependent kinase (CDK) inhibitor p27 and the transcription factors retinoic acid receptor β (RARβ) and Sox9. In mice bearing HNSCC PDX tumors, combined adjuvant and neo-adjuvant treatment with C26 resulted in complete inhibition of lethal lung metastasis. Mechanistic analysis showed that lung DTCs in C26-treated mice displayed an NR2F1hi/p27hi/Ki-67lo phenotype, which kept them dormant in a single-cell state preventing their outgrowth into overt metastases. Our work reveals a novel NR2F1 agonist and provides a proof of principle strategy supporting that inducing DTC dormancy using NR2F1 agonists could be used as a therapeutic strategy to prevent metastasis.

Current international guidelines recommend colonoscopy in patients with acromegaly at the time of diagnosis, even though the risk of developing colorectal neoplasm is still controversial. The main objective of this Argentine multicenter study was to analyze through screening colonoscopy the presence of advanced neoplastic lesions considered as precancerous, in patients with acromegaly compared to a control group. This is a case–control retrospective study. Full length colonoscopy of 70 acromegalic patients and 128 control subjects were studied. Polyps were classified into non pre-cancerous lesions and advance neoplastic lesions which included advanced adenomas (preneoplastic) and colorectal carcinomas. Thirty three out of 70 acromegalic patients and 32 out of 128 subjects controls presented polyps in the colonoscopy [47.1% vs 25%, p = 0.002, OR 2.68]. Non precancerous polyps were found in 11 (15.7%) and 23 (17.9%) (p = 0.690), while advanced neoplastic lesions were found in 22 (31.4%) and 9 (7.0%) (p = 0,0001 – OR: 6.06) patients and controls respectively. Advanced adenomas and colorectal carcinomas were found in 18 (27.3%) and 9 (7.0%) (p = 0,0006—OR: 4,57), and 4 (5.7%) and 0 (0.0%) p = 0.0063) of patients and controls respectively. The presence of insulin resistance was the only statistically significant associated factor among acromegalic patients with and without colonic polyps. Our findings show an increased risk of preneoplastic colonic lesions and colorectal carcinoma in patients with chronic and sustained GH excess compared to a control group. This supports the recommendation to perform screening colonoscopy at diagnosis of acromegaly.

Diagnosis of endogenous Cushing’s syndrome entails corticotropic autonomy, lack of circadian rhythm and/or hypercortisolism, evaluated through 24h urinary free cortisol (UFC). Hair cortisol measurement (HCM) has been described as an alternative marker of cortisol exposure over the preceding three months. OBJECTIVES To evaluate HCM in Cushing’s disease (CD). To analyze the correlation between HCM and UFC. To compare HCM values in CD vs controls. PATIENTS AND METHODS 3 cm hair from posterior vertex in CD and in controls age- and gender-matched between May 2017 and May 2019. Controls were low level stressed individuals (Holmes-Rahe’s scale) without adrenal disease. Normal reference interval of HCM was defined (40-128 pg/mg hair). Measurement: Siemens Immulite 2000 (Gwynedd, UK) automated chemoluminiscent immunoassay (CLIA) UFC values within the 3 months previous to hair collection were considered. Controlled CD defined as UFC ≤1 upper normal limit (UNL) with or without treatment, remission as UFC ≤1 without pituitary lesion. Results are presented as median (m) and range. Kruskal-Wallis ANOVA used for median difference evaluation and Kappa index for concordance determination. Chi2 test for comparison of recategorized UFC and HCM. Statistical analysis performed with SPSS 23.0 RESULTS 23 CD patients recruited, median age 42 ± 11 years; 91% (n=21) female; 10 samples collected at diagnosis and 13 during follow-up. Control group composed of 50 individuals 45% (n=10) had controlled CD (mUFC 0.42 UNL, range 0.1-0.9) and a mHCM of 134.5 pg/mg (62-334) and 55% (n=12) did not have control (mUFC 2.2, 1.1-6) and a mHCM of 150.5 (75-459). After recategorization of UFC (> o ≤ 1 UNL) and HCM (> o ≤ 128 pg/mg), determinations were associated (Chi2, p= 0.18), however, the concordance was acceptable (Kappa index = 0.276). After dividing CD patients according to HCM, 35% (n=8) had normal HCM: mHCM 113.5 (62-126) and mUFC 0.45 (0.1- 4.4). Among them, 63% (n=5) had controlled CD (mHCM 110, 62-121; mUFC 0.39, 0.1-0.85); 25% (n=2) had active CD (mUFC 2.7, 1.1-4.4; mHCM 121, 75-126). 65% had high HCM (n=15): mHCM 167 (132-459) and mUFC 1.36 (0.1-6). Most of them had active CD (n=11, 73%): mHCM 160 (132-459) and mUFC 2.2 (1.1-6). Four patients with elevated HCM (m 248, 148-334) had normal UFC (m 0.61, 0.12-0.92): 2 were in remission, 1 had normal postsurgical UFC with active disease in the follow-up and 1 had normal UFC under medical treatment. Controls (n=50) had mHCM 62.5 (40-126), significantly different from CD. CONCLUSIONS We evaluated HCM in CD, proposing this method as an additional diagnostic test for hypercortisolism. The acceptable concordance between UFC and HCM is possibly due to the different duration of the evaluated periods. The difference in the HCM values observed between controlled or active CD patients and controls permits the consideration of the method as an alternative in the diagnosis and/or follow-up of CD.

Disseminated tumor cells (DTCs) are recognized as the seeds of metastasis. However, metastatic lesions can become symptomatic years or decades after primary tumor removal. This clinical finding suggests that DTCs are not immediately competent to initiate growth and can persist in a dormant state. Here we review recent data for three potential scenarios that could result in DTC dormancy: (a) The target organ microenvironment instructs DTCs to enter dormancy; (b) primary tumors pre-encode a dormancy signature that only becomes evident when DTCs enter target organs that produce dormancy-inducing cues; and (c) early dissemination spawns DTCs that, by virtue of being closely related to normal cells, would retain the capacity to respond to dormancy-instructing signals and enter dormancy in target organs. The literature supports the existence of these scenarios and provides insight into how to prevent metastasis. Importantly, cotargeting dormant and proliferative DTCs in stage IV cancer may also improve outcomes in this clinical setting.

Spindle cell oncocytoma is an infrequent benign non-endocrine sellar neoplasm. Due to its similar morphology to pituitary adenomas, consideration of this differential diagnosis would conduce to a more careful surgical approach in order to avoid intraoperative bleeding and aiming to a complete resection, on which depends long-term outcomes. We present the case of a 60-year-old male who complained about visual abnormalities, with computerized visual field confirmation. On biochemistry, a panhypopituitarism was detected. The brain magnetic resonance images showed a sellar mass. A non-functioning pituitary macroadenoma was presumptively diagnosed and due to the visual impairment, surgical transesphenoidal treatment was indicated. The histological diagnosis was spindle cell oncocytoma. Five months after surgery, the control image demonstrated a lesion that was considered as remnant tumor, hence radiosurgery was performed. During the follow-up, the tumor reduced its size and four years after initial treatment, the sellar resonance imaging showed disappearance of the residual tumor. Communication of new cases of this rare entity will enlarge the existing evidence and will help to determinate the most effective treatment and prognosis.

Cancer cell dissemination during very early stages of breast cancer proceeds through poorly understood mechanisms. Here we show, in a mouse model of HER2+ breast cancer, that a previously described sub-population of early-evolved cancer cells requires macrophages for early dissemination. Depletion of macrophages specifically during pre-malignant stages reduces early dissemination and also results in reduced metastatic burden at end stages of cancer progression. Mechanistically, we show that, in pre-malignant lesions, CCL2 produced by cancer cells and myeloid cells attracts CD206+/Tie2+ macrophages and induces Wnt-1 upregulation that in turn downregulates E-cadherin junctions in the HER2+ early cancer cells. We also observe macrophage-containing tumor microenvironments of metastasis structures in the pre-malignant lesions that can operate as portals for intravasation. These data support a causal role for macrophages in early dissemination that affects long-term metastasis development much later in cancer progression. A pilot analysis on human specimens revealed intra-epithelial macrophages and loss of E-cadherin junctions in ductal carcinoma in situ, supporting a potential clinical relevance., Early dissemination of cancer cells has been reported to occur in certain breast cancer models. Here the authors show that intra-epithelial macrophages in the early pre-cancer lesions drive early cancer cell dissemination through Wnt-1 secretion and that such events impact the later development of metastasis.

SUMMARY Chordoid glioma (CG) is considered a slow growing glial neoplasm. We report two new cases with endocrinological presentation, management and outcome. Case reports: 1) An 18 year-old female patient was admitted due to headaches, nausea and vomiting and visual abnormalities. She was in amenorrhea. A brain magnetic resonance imaging (MRI) demonstrated a 35 mm-diameter sellar and suprasellar mass. An emergency ventricular peritoneal valve was placed due to obstructive hydrocephalus. Transcraneal surgery was performed. The patient developed central hypothyroidism, adrenal insufficiency and transient diabetes insipidus; she never recovered spontaneous menstrual cycles. Histopathologic study showed cells in cords, inside a mucinous stroma, positive for glial fibrillary acidic protein (GFAP). Due to residual tumor gamma knife radiosurgery was performed. Three years after surgery, the patient is lucid, with hypopituitarism under replacement. 2) A 46 year-old woman complained about a three year-history of amenorrhea, galactorrhea and headache. An MRI showed a solid-cystic sellar mass 40 mm-diameter that extended to the suprasellar cistern. She had hypogonatropic hypogonadism and mild hyperprolactinemia. The tumor mass was removed via nasal endoscopic approach. Histopathological study reported cellular proliferation of glial lineage positive for GFAP. The patient evolved with central hypothyroidism and diabetes insipidus. She was re-operated for fistula and again under the diagnosis of extradural abscess. She evolved with cardiorespiratory descompensation and death, suspected to be due to a thromboembolism. In conclusion, the first case confirms that best treatment for CG is surgery considering radiotherapy as an adjuvant therapy. The other case, on the contrary, illustrates the potentially fatal evolution due to surgical complications.

Cancer cells disseminate during very early and sometimes asymptomatic stages of tumor progression. Granted that biological barriers to tumorigenesis exist, there must also be limiting steps to early dissemination, all of which remain largely unknown. We report that the orphan nuclear receptor NR2F1/COUP-TF1 serves as a barrier to early dissemination. High-resolution intravital imaging revealed that loss of function of NR2F1 in HER2+ early cancer cells increased in vivo dissemination without accelerating mammary tumor formation. NR2F1 expression was positively regulated by the tumor suppressive MMK3/6-p38-MAPK pathway and downregulated by HER2 and Wnt4 oncogenic signaling. NR2F1 downregulation by HER2 in early cancer cells led to decreased E-cadherin expression and β-catenin membrane localization, disorganized laminin 5 deposition, and increased expression of CK14, TWIST1, ZEB1 and PRRX1. Our findings reveal the existence of an inhibitory mechanism of dissemination regulated by NR2F1 downstream of HER2 signaling. Citation Format: Maria Soledad Sosa, Carolina Rodriguez-Tirado, Nupura Kale, Maria Carlini, Nitisha Shrivastava, Bassem Khalil, Javier Bravo-Cordero, Melissa Alexander, Jiayi Ji. NR2F1 is a barrier to early dissemination of pre-malignant mammary cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2837.

Resumen Introduccion El riesgo de desarrollar neoplasias colonicas en pacientes acromegalicos y su relacion directa con los niveles elevados de GH/IGF-1 no estan bien establecidos y continuan siendo motivo de controversia en la literatura mundial. El objetivo de este trabajo fue evaluar el riesgo de desarrollar lesiones neoplasicas avanzadas (LNA) (adenomas mayores a 1 cm, componente velloso mayor del 75% y/o displasia de alto grado), en pacientes con acromegalia, comparado con un grupo control. Materiales y metodos Estudio multicentrico caso-control retrospectivo. Ciento treinta y siete pacientes con acromegalia que realizaron videocolonoscopia (VCC) fueron incluidos inicialmente, aunque solo 69 cumplieron criterios de inclusion. Sesenta y dos controles fueron obtenidos: por cada caso (paciente con acromegalia) 2 «controles» fueron seleccionados aleatorizadamente e igualados por edad y sexo. El riesgo se expreso en odds ratio (OR) y su correspondiente intervalo de confianza (IC) del 95%. La significacion estadistica fue considerada una p Resultados De los 69 pacientes con VCC completa y datos adecuados para su analisis, 28 presentaron VCC positiva con hallazgos de polipos (40%) y 41 VCC negativa o normal (60%). Dentro del grupo con VCC positiva, 14 presentaron LNA (20%) y solo un paciente presento diagnostico de cancer colorrectal. Para el analisis caso-control se incluyo a 31 pacientes frente al grupo control (n = 62) que cumplieron con los criterios de inclusion. La presencia de polipos colonicos, adenomas y LNA en los pacientes con acromegalia fue de 19/31 (61,9%), 14/31 (45,16%) y 10/31 (32,25%), y en el grupo control de 18/62 (29,03%), 11/62 (17,74%) y 4/62 (6,45%), respectivamente. El riesgo de adenomas y LNA fue mayor en el grupo de acromegalia en comparacion con el grupo control, siendo ambos resultados estadisticamente significativos: adenomas OR 2,54 (IC 1,22-5,25) p = 0,005, LNA OR: 7,3 (2,4-25), p = 0,00. Conclusion La acromegalia se asocia a un mayor riesgo de lesiones colonicas preneoplasicas. Este hallazgo justifica el cribado con VCC al diagnostico en pacientes con acromegalia.

WHO (2012) reports that chronic exposure to air pollutants, including particulate matter (PM), causes the death of 7 million people, constituting the most important environmental risk for health in the world. IARC classifies contaminated outdoor air as carcinogenic, Group 1 category. However, in our countries there are few studies regarding air pollution levels and possible associated effects on public health. The current study determined PM and associated polycyclic aromatic hydrocarbons (PAHs) levels in outdoor air, identified their possible emission sources and analysed health risks in the city of Tandil (Argentina). PM10 and PM2.5 samples were collected using a low volume sampler (MiniVol TAS) in three areas: city centre, industrial and residential. Concentrations were determined by gravimetric methods and the content of the US EPA 16 priority PAHs was found by high performance liquid chromatography (HPLC). Description of the main emission sources and selection of monitoring sites resulted from spatial analysis and the IVE (International Vehicle Emissions) model was used in the characterisation of the traffic flow. Median values of 35.7 μgm-3 and 9.6 μgm-3 in PM10 and PM2.5 respectively and characteristic profiles were found for each area. Local values PAHs associated to PM10 and PM2.5, in general, were lower than 10ngm-3. The estimated Unit Risk for the three areas exceeds US EPA standards (9 × 10−5). The number of deaths attributable to short term exposure to outdoor PM10 was 4 cases in children under 5 years of age, and 21 cases in total population, for a relative risk of 1.037. Fil: Sosa, Beatriz Soledad. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Humanas. Departamento de Ciencias Ambientales. Centro de Investigación y Estudios Ambientales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Porta, Atilio Andrés. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Química. Centro de Investigaciones del Medio Ambiente; Argentina Fil: Colman Lerner, Jorge Esteban. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Química. Centro de Investigaciones del Medio Ambiente; Argentina Fil: Banda Noriega, Roxana Bettina. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Humanas. Departamento de Ciencias Ambientales. Centro de Investigación y Estudios Ambientales; Argentina. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas; Argentina Fil: Massolo, Laura Andrea. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Química. Centro de Investigaciones del Medio Ambiente; Argentina

Pathologists rely on microscopy to diagnose disease states in tissues and organs. They utilize both high-resolution, high-magnification images to interpret the staining and morphology of individual cells, as well as low-magnification overviews to give context and location to these cells. Intravital imaging is a powerful technique for studying cells and tissues in their native, live environment and can yield sub-cellular resolution images similar to those used by pathologists. However, technical limitations prevent the straightforward acquisition of low-magnification images during intravital imaging, and they are hence not typically captured. The serial acquisition, mosaicking, and stitching together of many high-resolution, high-magnification fields of view is a technique that overcomes these limitations in fixed and ex vivo tissues. The technique however, has not to date been widely applied to intravital imaging as movements caused by the living animal induce image distortions that are difficult to compensate for computationally. To address this, we have developed techniques for the stabilization of numerous tissues, including extremely compliant tissues, that have traditionally been extremely difficult to image. We present a novel combination of these stabilization techniques with mosaicked and stitched intravital imaging, resulting in a process we call Large-Volume High-Resolution Intravital Imaging (LVHR-IVI). The techniques we present are validated and make large volume intravital imaging accessible to any lab with a multiphoton microscope.

Metastasis is the leading cause of cancer-related deaths; metastatic lesions develop from disseminated cancer cells (DCCs) that can remain dormant. Metastasis-initiating cells are thought to originate from a subpopulation present in progressed, invasive tumours. However, DCCs detected in patients before the manifestation of breast-cancer metastasis contain fewer genetic abnormalities than primary tumours or than DCCs from patients with metastases. These findings, and those in pancreatic cancer and melanoma models, indicate that dissemination might occur during the early stages of tumour evolution. However, the mechanisms that might allow early disseminated cancer cells (eDCCs) to complete all steps of metastasis are unknown. Here we show that, in early lesions in mice and before any apparent primary tumour masses are detected, there is a sub-population of Her2+p-p38lop-Atf2loTwist1hiE-cadlo early cancer cells that is invasive and can spread to target organs. Intra-vital imaging and organoid studies of early lesions showed that Her2+ eDCC precursors invaded locally, intravasated and lodged in target organs. Her2+ eDCCs activated a Wnt-dependent epithelial-mesenchymal transition (EMT)-like dissemination program but without complete loss of the epithelial phenotype, which was reversed by Her2 or Wnt inhibition. Notably, although the majority of eDCCs were Twist1hiE-cadlo and dormant, they eventually initiated metastasis. Our work identifies a mechanism for early dissemination in which Her2 aberrantly activates a program similar to mammary ductal branching that generates eDCCs that are capable of forming metastasis after a dormancy phase.

Al abordar la problemática de la contaminación del aire, se sabe que son muy variadas las actividades que pueden resultar en fuentes de contaminación y contribuir (con diferente magnitud) a esta situación. En general, los esfuerzos por proteger este recurso se dirigen en dos direcciones: los que buscan evitar problemas consecuentes de la contaminación en el largo plazo (ej. cambio climático, efecto invernadero), y los que buscan dar soluciones a efectos nocivos de los contaminantes en la salud humana en el corto y mediano plazo. En este contexto, el deterioro de la calidad del aire representa una amenaza importante para la salud pública en todo el mundo. Según la Organización Mundial de la Salud (OMS) más de dos millones de muertes prematuras anuales son atribuibles a los efectos de la contaminación del aire en espacios abiertos urbanos y en espacios cerrados. Las actividades que se desarrollan en el ambiente urbano no quedan ajenas a ello incluso, en algunos casos, tienen el agravante de condiciones meteorológicas desfavorables que acentúan la persistencia de los contaminantes en la atmósfera local. Ejemplos de ello se evidencian claramente en grandes urbes como ciudad de México, Santiago de Chile, Los Ángeles, Nueva Deli, Buenos Aires, entre otras; con el problema ambiental ya reconocido por sus sociedades. Si abordamos la calidad del aire en ciudades chicas o intermedias aparece, en general, la dificultad de la falta de recursos técnicos y humanos que permitan conocer la situación local. La ciudad de Tandil (República Argentina), en relación a su entorno, aparece como un municipio con síntomas típicos del crecimiento de un centro urbano medio-grande, donde la pérdida de la calidad del aire como consecuencia de la actividad industrial de fundición, por la descarga de efluentes gaseosos dentro del casco urbano, y el rápido crecimiento del parque automotor son muestra de ello. Es por ello que el presente trabajo de tesis doctoral tiene por objeto caracterizar la calidad del aire (en particular compuestos orgánicos volátiles, PM10 y PM2,5 y compuestos adsorbidos) en la ciudad bonaerense de Tandil, identificar y evaluar el riesgo asociado a la salud, y elaborar indicadores proponiendo una red de monitoreo que permita evaluar la problemática en cuestión y su evolución en el tiempo a fin de brindar herramientas para la toma de decisiones. La caracterización de los compuestos presentes en calidad de aire se efectúa mediante muestreos estacionales, en campañas, para el periodo 12/2010 a 07/2013 para COV, y 03/2013 a 09/2014 para PM. Las observaciones se hacen en tres zonas de características urbanas diferentes industrial / centro / control. La captación de partículas inhalables (PM10) y respirables (PM 2,5) se realiza mediante muestreador portátil de bajo volumen (MiniVol Tas) en un periodo de 5 días por muestra; se realizan en ellas análisis gravimétrico, espectrofotometría de absorción atómica para la determinación de metales y, cromatografía líquida de alta resolución con detección de fluorescencia para la determinación de HAP. En la caracterización de COV, las muestras se toman en un periodo de 30 días mediante captador pasivo 3M 3500; el análisis se efectúa mediante cromatografía gaseosa. La estimación del riesgo asociado a compuestos de interés para la salud se logra mediante la utilización de la metodología sugerida por la OMS/ USEPA. Las fuentes principales de emisiones a la atmósfera se describen y caracterizan a partir del relevamiento de información en campo; se realizan encuestas a informantes calificados para el conocimiento de las condiciones operativas de las industrias de fundición. En paralelo, la caracterización del parque automotor, se desarrolla con el aporte de la base de datos del Ente Regulador de la verificación Técnica Vehicular la cual se analiza mediante estadística descriptiva y la utilización del software Infostat; este trabajo se complementa además con el relevamiento del flujo vehicular en las tres zonas mencionadas. Las variaciones de las concentraciones halladas en PM10 y PM2,5, así como valores identificados para sus compuestos asociados (metales y HAP) señalan eventos que afectan y condicionan el análisis de los resultados, impidiendo hacer generalizaciones o encontrar patrones. Lo resultados de las observaciones para HAP son dispersos e implican la necesidad de seguir repitiendo los muestreos a fin de conocer en qué medida las explicaciones encontradas para los valores altos -de algunos compuestos- se corresponden o no con situaciones puntuales. Por otra parte, la carencia de diferencias evidentes en la presencia/ausencia de metales esperables en las distintas zonas estudiadas (industrial, centro y control), se atribuye a las características de los suelos por el uso urbano (tanto presente como pasado). Respecto de los COV, se dieron variaciones de importancia a lo largo de las campañas al comparar entre zonas, y también entre campaña y campaña para un mismo sitio, sobre todo cuando se observa el ciclohexano, el cloroformo, el benceno y tolueno; si bien el tolueno se encuentra dentro de los valores guías establecidos por la legislación, el benceno supera en todas las zonas dicho límite. Los resultados indican riesgo individual de contraer cáncer (RICV) por benceno a lo largo de la vida en todas las zonas estudiadas en el orden de 1 x 10-5, aunque el mayor RICV se da en la zona centro, y el menor en la zona control. Por otra parte, la ∑ BaP eq. de HAP en PM10 y PM2,5 muestra un incremento en el riesgo individual de contraer cáncer que excede lo establecido por la USEPA de 1 x 10-6, pero que se encuentran cercanos al límite de 9 x 10-5 que establece la OMS. Existen además en el ambiente urbano otros compuestos preocupantes para la salud de la población de Tandil, como son cloruro de carbono / cloroformo / plomo, que se categorizan como probablemente carcinogénicos (Grupo 2B). Es necesario dar continuidad a la generación de datos que informen sobre la calidad de aire local para tener un conocimiento más certero de la relación que existe entre las fuentes y los valores hallados en inmisión. De todos modos, se pudo concluir que los resultados alcanzados, junto a la caracterización de las fuentes emisoras principales, aportan un diagnóstico de la situación actual. Este diagnóstico es un punto de partida importante para la toma de decisiones del gobierno local -y su gestión ambiental- respecto del recurso aire, debido a que es el primer estudio de estas características que se realiza en la ciudad. De acuerdo con lo expresado, la batería de indicadores propuestos (2 para concentraciones de PM; 7 para metales adsorbidos a material particulado; 7 para HAP adsorbidos en PM; y, 6 para COV), junto al establecimiento de la red de monitoreo elaborada para las condiciones particulares de este área de estudio, resultan en una herramienta clave para planificar propuestas de manejo sustentables del recurso., Facultad de Ciencias Exactas

Introduction: Pituitary Metastases (MTS) are infrequently seen in clinical practice. The incidence ranges from 0.14 to 28%. Breast and lung cancer are the primary sites that most frequently metastasize to sellar region, between the sixth and seventh decades of life. Most cases are diagnosed in patients with advanced malignant disease, however, in 20-30%, symptoms of pituitary involvement can precede the diagnosis of the primary tumor. Objectives: To evaluate symptoms at presentation, hormonal, radiological and histological findings, management and outcome of a series of patients with pituitary MTS. Patients and methods: medical records of 16 patients from eight Endocrine Centers were reviewed. Ten patients had histological confirmation of the pituitary MTS, 6 were not operated, being the diagnostic criteria the presence of sellar mass associated with diabetes insipidus (DI) and / or sudden-onset of neuro-ophthalmological symptoms in patients with confirmed primary neoplasia. Results: The median age was 54 years (range 35-70), 9 women (56.2%). The sites of the primary tumor were: 7 lung (44%), 5 breast (32%), 1 follicular thyroid carcinoma (6%), 1 Hodgkin lymphoma (6%), 1 poorly differentiated carcinoma (6%), and 1 clear cell renal carcinoma (6%). The median time between the diagnosis of the primary neoplasm and the occurrence of the pituitary MTS was 12 months (range: 3-120). In 9 patients (56.2%), the diagnosis of the primary neoplasm was made after the finding of the symptomatic sellar mass. DI was found in 14 patients (87.5%), adenohypophyseal deficit in 12 (75%), visual disorders in 10 (62.5 %), headache in 6 (37.5%) and cranial nerve deficits in 6 (37.5%). In 68.7% (11 patients), other MTS were detected. Fifteen patients were evaluated by MRI and one by CT: 13 (81.3%) harbored supra / parasellar masses, and the remaining 3 had lesions limited to the pituitary gland, with stalk thickening and lack of spontaneous neurohypophysis hyperintensity in 2 of them. In all cases diffuse gadolinium uptake was present. Fourteen patients died (87.5%), with a median survival time of 6,5 months (range: 1-36); the remaining 2 are still alive with a follow-up period of 4 and 12 months respectively. Conclusions: In this series of 16 patients with pituitary MTS, the most frequent primary neoplasms were lung and breast. Median age was lower than in published series. DI was the most common condition at presentation, followed by hypopituitarism and visual disorders. The short survival was related to the extent of the disease at the time of diagnosis. In more than half of the cases the diagnosis of primary neoplasia was made through the symptomatic pituitary mass. In the presence of a pituitary lesion with diffuse gadolinium uptake, associated with DI and / or acute visual deficit, pituitary MTS should be suspected even in patients without a history of oncological disease.

Prolactinomas represent approximately 40-60% of all pituitary tumors. Dopamine agonists (DA) are the first-line treatment, with a 70-90% response. However, up to 10% are resistant (R) to DA. Our objectives were to describe the clinical, biochemical and imaging characteristics of a group of patients with prolactinomas R to DA; and to determine the presence of prognostic factors by performing a comparative analysis with the population of non-R prolactinomas. We analized retrospectively the medical records of patients diagnosed with prolactinoma, with follow-up (f-up) at the same hospital from 1998 to 2017. They were classified as R when prolactin (PRL) was not normalized and/or a reduction ≤ than 50% of the tumor (Tx) volumen, with a cabergoline (CBG) dose ≥2mg/week (mg/w), with, at least 6 months of treatment. Macroadenomas (MA) were defined as Tx ≥10mm and as invasive with a classification of Knosp 3 or 4 by MRI. Of 95 prolactinomas, 14 patients with R to DA were described. The mean age was 32.7 ± 15.6 years (y). We found no statistically significant differences (SSD) in terms of gender, with a higher distribution (53%) and a tendency to age (37.9 vs 26.9, p = 0.09) in the male gender (M). The mean time of f-up was 34.7 ± 28.7 months (m). Majority were MA (87%) and invasive Tx (64%). The initial manifestations were: headache (85.7%), visual field alteration (50%), menstrual cycle alterations (86%), galactorrhea (20%). The mean PRL value was 7419.4 ± 1149.7 ng/ml, being higher in the M group (11652.6 vs 1653.3, p

Our understanding of the minimal residual disease (MRD) in solid cancers indicates that it can persist in the system for years or even decades. We now know that the persistence of MRD might depend on the dormancy of the disseminated cancer cells (DCCs). Once DCCs exit dormancy, they become metastatic and the survival rates of the patients inevitably decrease. Thus, innovative treatments are required to extend the asymptomatic phase of MRD after the initial therapeutic intervention. With the latest advances in cancer research, there is a greater need to explore and understand the biology, timing of dissemination, and origin of DCCs during tumor progression. These important aspects of DCCs impact the selection, design, administration, and timing of effective therapies. Herein, we summarize the current understanding of MRD biology in solid tumors, with a focus on epigenetics and pluripotency, presenting an overall view of the direction the field is taking to reach the goal of reducing cancer-related mortalities that result from metastasis.

Cancer cells have been found to disseminate during very early and sometimes asymptomatic stages of tumor progression. At secondary organs, early disseminated cancer cells (eDCCs) can enter a non-proliferative state for prolonged periods of time. Intriguingly, eDCCs also carry metastasis- initiating capacity. However, the mechanisms by which these early cancer cells complete all steps of metastasis including metastasis-initiating capacity are unknown. Here we report that the orphan nuclear receptor and lineage commitment regulator NR2F1 serves as an early barrier to early dissemination propelled by HER2 signaling. NR2F1 loss of function in HER2 early cancer cells induced cell motility, loss of E-cadherin junctions, disorganized laminin-V deposition, decreased β-catenin membrane localization and increased micro-invading cells with CK14+/CK18 phenotype. Importantly, upon NR2F1 depletion in HER2 early cancer acini, TWIST, ZEB1 and PRRX1 levels were upregulated. Our findings reveal a previously unrecognized function of NR2F1 as a suppressor of dissemination during early stages of breast tumor progression. Thus, therapies that restore and activate NR2F1 function might limit early dissemination and metastatic outgrowth. Citation Format: Maria Soledad Sosa, Nupura Kale, Nitisha Shrivastava, Carolina Rodriguez-Tirado, Maria Carlini, Jiayi Ji, Javier Bravo-Cordero. NR2F1 limits dissemination of early cancer mammary epithelial cells [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6061.

A method for the simultaneous determination of three commonly used coccidiostats in chicken liver was developed, comprising a multi-residue QuEChERS (quick, easy, cheap, effective, rugged and safe) extraction step, and a liquid chromatography-ultra violet-fluorescence (HPLC-UV/FL) analysis. The QuEChERS extraction was optimized using an experimental design approach that includes a screening step to obtain the critical variables, an optimization step using multiple response surface analysis and the calculation of a desirability parameter. The optimized method was validated with fortified samples, reaching an average recovery of 91% and an overall precision of 5.5% (mean of three analytes at three levels). Limits of detection calculated on fortified samples were 20 µg kg−1 for lasalocid, 15 µg kg−1 for nicarbazin and 120 µg kg−1 for diclazuril. These values resulted at least one order of magnitude lower than the maximum allowed residue limit (MRL) of the studied coccidiostats for chicken liver.

Background The presence of disseminated tumor cells (DTCs) in bone marrow (BM) is an independent prognostic factor in early breast cancer but does not uniformly predict outcome. Tumor cells can persist in a quiescent state over time, but clinical studies of markers predicting the awakening potential of DTCs are lacking. Recently, experiments have shown that NR2F1 (COUP-TF1) plays a key role in dormancy signaling. Methods We analyzed the NR2F1 expression in DTCs by double immunofluorescence (DIF) staining of extra cytospins prepared from 114 BM samples from 86 selected DTC-positive breast cancer patients. Samples collected at two or more time points were available for 24 patients. Fifteen samples were also analyzed for the proliferation marker Ki67. Results Of the patients with detectable DTCs by DIF, 27% had ≥ 50% NR2F1high DTCs, chosen a priori as the cut-off for “dormant profile” classification. All patients with systemic relapse within 12 months after BM aspiration carried ≤ 1% NR2F1high DTCs, including patients who transitioned from having NR2F1high-expressing DTCs in previous BM samples. Of the patients with serial samples, half of those with no relapse at follow-up had ≥ 50% NR2F1high DTCs in the last BM aspiration analyzed. Among the 18 relapse-free patients at the time of the last DTC-positive BM aspiration with no subsequent BM analysis performed, distant disease-free intervals were favorable for patients carrying ≥ 50% NR2F1high DTCs compared with those with predominantly NR2F1low DTCs (p = 0.007, log-rank). No survival difference was observed by classification according to Ki67-expressing DTCs (p = 0.520). Conclusions Our study translates findings from basic biological analysis of DTC dormancy to the clinical situation and supports further clinical studies of NR2F1 as a marker of dormancy.

Primary hypophysitis (PH) is a rare disease with a poorly-defined natural history. Our aim was to characterise patients with PH at presentation and during prolonged follow-up. Observational retrospective study of 22 patients was conducted from 3 centres. In 14 patients, PH was confirmed histologically and in the remaining 8 clinically, after excluding secondary causes of hypophysitis. All patients had hormonal and imaging investigations before any treatment. Median follow up was 48 months (25–75%: 3–60). There was a female predominance with a female/male ratio: 3.4:1. Eight out of 22 patients had another autoimmune disease. Headaches and gonadal dysfunction were the most common symptoms. Five patients presented with panhypopituitarism; 17 patients had anterior pituitary deficiency, and 7 had diabetes insipidus. At presentation, 9 patients were treated surgically, 5 received replacement hormonal treatment, and 8 high-dose glucocorticoids from whom 5 in association with other immunosuppressive agents. Six patients showed complete recovery of pituitary hormonal deficiencies while 6 showed a partial recovery during a 5-year follow-up period. No difference was found between patients treated with surgery and those treated medically. The overall relapse rate was 18%. PH can be manifested with a broad spectrum of clinical and hormonal disturbances. Long-term follow-up is required to define the natural history of the disease and response to treatment, since pituitary hormonal recovery or relapse may appear many years after initial diagnosis. We suggest that surgery and immunosuppressive therapy be reserved for exceptional cases.

Our understanding of the minimal residual disease (MRD) in solid cancers indicates that it can persist in the system for years or even decades. We now know that the persistence of MRD might depend on the dormancy of the disseminated cancer cells (DCCs). Once DCCs exit dormancy, they become metastatic and the survival rates of the patients inevitably decrease. Thus, innovative treatments are required to extend the asymptomatic phase of MRD after the initial therapeutic intervention. With the latest advances in cancer research, there is a greater need to explore and understand the biology, timing of dissemination, and origin of DCCs during tumor progression. These important aspects of DCCs impact the selection, design, administration, and timing of effective therapies. Herein, we summarize the current understanding of MRD biology in solid tumors, with a focus on epigenetics and pluripotency, presenting an overall view of the direction the field is taking to reach the goal of reducing cancer-related mortalities that result from metastasis.

Metastasis is responsible for the vast majority of cancer-related deaths. However, our understanding of this complex process is still vastly limited, as is the ability to prevent metastasis. Paradoxically, while clinical trials are commonly performed in patients with advanced metastatic disease, disseminated residual disease is rarely targeted. This eliminates a critical window of opportunity to prevent metastasis. Disseminated tumor cells (DTCs) that seed metastases can remain undetected years to decades after treatment of the primary tumor. Late relapse may be due to the ability of DTCs to survive in a quiescent or dormant state and evade therapies. Quiescence, a reversible growth arrest coupled to robust survival, has emerged as a fitting biological definition for dormancy of single DTCs, but these mechanisms remain as one of the least understood “black boxes” in cancer biology. Because of the reversible nature of dormancy, it has been proposed that epigenetic changes are key in regulating the onset, maintenance and reactivation from this state. This is mediated by the post-translational modification of histones (PTMs), ATP-dependent chromatin remodeling, DNA methylation, and the incorporation of specialized histone variants into chromatin. Many morphogenetic and micro-environmental cues like retinoic acid, TGFβs, hematopoietic stem cell dormancy regulating cues and BMPs are known to cause chromatin modifications that dictate cell fate; these same cues were linked to the induction of cancer cell dormancy. Despite progress in understanding cancer cell dormancy, key questions remain open regarding its epigenetic nature. In this chapter we attempt to address key questions related to this topic using available data or hypothetical scenarios to build a model to further dissect how cancer cell dormancy can be manipulated epigenetically as a therapeutic strategy.

Nowadays, environmental indicators are widely used as effective tools to assist decision-making in both public and private sectors. The lack of literature and research about local and regional Environmental Condition Indicators (ECI), the poor knowledge regarding solid waste generation, effluents and gas emissions from foundry industries, and their particular location in the urban area of Tandil, Argentina are the main reasons for this investigation, aiming to develop a set a of ECI to provide information about the environment in relation to the foundry industry. The study involves all the foundries located in the city between March and April 2010. The set of ECI developed includes 9 indicators for air, 5 for soil and 1 for water. Specific methodology was used for each indicator.El empleo de indicadores ambientales es, actualmente, una herramienta que colabora en el proceso de toma de decisiones en la gestión de la administración pública y privada. La falta de antecedentes de Indicadores de Condición Ambiental (ICA) en lo regional y local, la información asociada a la generación de residuos, efluentes y emisiones gaseosas provenientes de la industria de fundición, y la particularidad de encontrar a estas industrias en el ámbito urbano en la ciudad de Tandil, Argentina, orientan el principal objetivo del presente trabajo: la construcción de un conjunto de ICA que proporcionen información sobre el estado del ambiente en relación a este tipo de actividad. El campo de la investigación comprendió el total de industrias de fundición de la ciudad en el periodo marzo – abril de 2010. Se generaron ICA para el recurso, aire (9), suelo (5) y agua (1) con metodología adaptada a cada caso particular.

Accumulating data suggest that metastatic dissemination often occurs early during tumour formation, but the mechanisms of early metastatic spread have not yet been addressed. Here, by studying metastasis in a HER2-driven mouse breast cancer model, we show that progesterone-induced signalling triggers migration of cancer cells from early lesions shortly after HER2 activation, but promotes proliferation in advanced primary tumour cells. The switch from migration to proliferation was regulated by increased HER2 expression and tumour-cell density involving microRNA-mediated progesterone receptor downregulation, and was reversible. Cells from early, low-density lesions displayed more stemness features, migrated more and founded more metastases than cells from dense, advanced tumours. Notably, we found that at least 80% of metastases were derived from early disseminated cancer cells. Karyotypic and phenotypic analysis of human disseminated cancer cells and primary tumours corroborated the relevance of these findings for human metastatic dissemination.

Rac GTPases, small G-proteins widely implicated in tumorigenesis and metastasis, transduce signals from tyrosine-kinase, G-protein-coupled receptors (GPCRs), and integrins, and control a number of essential cellular functions including motility, adhesion, and proliferation. Deregulation of Rac signaling in cancer is generally a consequence of enhanced upstream inputs from tyrosine-kinase receptors, PI3K or Guanine nucleotide Exchange Factors (GEFs), or reduced Rac inactivation by GTPase Activating Proteins (GAPs). In breast cancer cells Rac1 is a downstream effector of ErbB receptors and mediates migratory responses by ErbB1/EGFR ligands such as EGF or TGFα and ErbB3 ligands such as heregulins. Recent advances in the field led to the identification of the Rac-GEF P-Rex1 as an essential mediator of Rac1 responses in breast cancer cells. P-Rex1 is activated by the PI3K product PIP3 and Gβγ subunits, and integrates signals from ErbB receptors and GPCRs. Most notably, P-Rex1 is highly overexpressed in human luminal breast tumors, particularly those expressing ErbB2 and estrogen receptor (ER). The P-Rex1/Rac signaling pathway may represent an attractive target for breast cancer therapy.