Evaluación de la expresión de PD-L1 e infiltración linfocitaria intratumoral en carcinoma de mama invasivo triple negativo / Evaluation of PD-L1 expression and intratumoral lym phocytic infiltration in triple-negative invasive breast car cinoma

Resumen Introducción : El endofenotipo de cáncer de mama triple negativo (TNBC) es uno de los menos frecuentes y sin diana terapéutica, por tanto, se plantea estudiar la correlación del punto de control inmunológico PD-L1 con el establecimiento de microambiente tumoral evaluado por la infiltración linfocitaria intratumoral estromal (TILs) y su importancia en la práctica clínica. Métodos : Se realizó un estudio retrospectivo de casos y controles, con 31 casos de carcinoma infiltrante de la mama triple negativo y 57 controles no pareados de endofenotipo Luminal A, Luminal B y HER-2 atendidos en un año. Se evaluaron las variables: tipo y grado his tológico, expresión PD-L1 con el clon 22C3, TILs, invasión linfovascular, tamaño tumoral, compromiso de ganglios linfáticos y metástasis. El análisis estadístico se ejecutó con la prueba de chi cuadrado y prueba de coeficiente de correlación de Spearman. Resultados : Se encontró una correlación negativa estadísticamente significativa entre TILs y PD-L1 (rho - 0.106, p 0.025), indicando que a mayor expresión de PD-L1, es menor la infiltración linfocitaria intratumo ral. En los grupos de TILs B (10-40% TILs) y C (40-90% TILs) donde se presenta marcado infiltrado inflamatorio intratumoral se evidenció mayor número de pacientes negativos para PD-L1 (CPS <10) con 16 y 10 casos res pectivamente. Para los casos TNBC se logró identificar un coeficiente de asociación negativa (rho -0.378) y con significancia estadística (p 0.01). Discusión : Se estableció la asociación de TNBC, TILs y expresión de PDL1, lo cual es importante para la instau ración de terapias diana y el desarrollo de la medicina de precisión.

Abstract Introduction : Triple negative breast cancer endophe notype (TNBC) is one of the least frequent and without therapeutic target; therefore we propose to study the correlation of PD-L1 immune checkpoint with the es tablishment of tumor microenvironment assessed by intratumoral stromal lymphocyte infiltration (TILS) and its importance in clinical practice. Methods : A retrospective case-control study was performed, with 31 cases of triple-negative infiltrat ing breast carcinoma and 57 unmatched controls of Luminal A, Luminal B and HER-2 endophenotype seen in one year. The following variables were evaluated: histologic type and grade, PD-L1 expression with clone 22C3, TILS, lymphovascular invasion, tumor size, lymph node involvement and metastasis. Statistical analysis was performed with the chi-square test and Spearman correlation coefficient test. Results : a statistically significant negative correlation was found between TILS and PD-L1 (rho - 0.106, p 0.025), indicating that the higher the expression of PD-L1, the lower the intratumoral lymphocytic infiltration. In the TILS B (10-40% TILS) and C (40-90% TILS) groups where there was a marked intratumoral inflammatory infiltrate, a greater number of patients were negative for PD-L1 (CPS <10) with 16 and 10 cases, respectively. For TNBC cases a negative association coefficient was identified (rho -0.378) with statistical significance (p 0.01). Discussion : The association between TNBC, TILS and PDL1 expression was established, which is important for the establishment of target therapies and the develop ment of precision medicine.

Machine learning applications on intratumoral heterogeneity in glioblastoma using single-cell RNA sequencing data.

Artificial intelligence is revolutionizing all fields that affect people's lives and health. One of the most critical applications is in the study of tumors. It is the case of glioblastoma (GBM) that has behaviors that need to be understood to develop effective therapies. Due to advances in single-cell RNA sequencing (scRNA-seq), it is possible to understand the cellular and molecular heterogeneity in the GBM. Given that there are different cell groups in these tumors, there is a need to apply Machine Learning (ML) algorithms. It will allow extracting information to understand how cancer changes and broaden the search for effective treatments. We proposed multiple comparisons of ML algorithms to classify cell groups based on the GBM scRNA-seq data. This broad comparison spectrum can show the scientific-medical community which models can achieve the best performance in this task. In this work are classified the following cell groups: Tumor Core (TC), Tumor Periphery (TP) and Normal Periphery (NP), in binary and multi-class scenarios. This work presents the biomarker candidates found for the models with the best results. The analyses presented here allow us to verify the biomarker candidates to understand the genetic characteristics of GBM, which may be affected by a suitable identification of GBM heterogeneity. This work obtained for the four scenarios covered cross-validation results of $93.03\% \pm 5.37\%$, $97.42\% \pm 3.94\%$, $98.27\% \pm 1.81\%$ and $93.04\% \pm 6.88\%$ for the classification of TP versus TC, TP versus NP, NP versus TP and TC (TPC) and NP versus TP versus TC, respectively.

Intratumoral Treatment with 5-Androstene-3ß, 17α-Diol Reduces Tumor Size and Lung Metastasis in a Triple-Negative Experimental Model of Breast Cancer.

Breast cancer treatment failure is related to low response rates, high costs, and long-term toxicities. Thus, it is necessary to find less toxic, cheaper, and more effective treatments. In situ administration ensures drug delivery to tumor cells and decreases systemic toxic effects. The androstene-3ß, 17α-diol (α-AED) reduces breast tumor cell proliferation and is an ideal candidate to treat mammary tumors. This study aims to identify the in vitro and in vivo effects of α-AED on a triple-negative mammary tumor model. An in vitro biphasic steroid effect was observed in mouse and human mammary tumor cells treated with α-AED. In this sense, cells treated with higher doses (100 and 200 µM) showed an antiproliferative effect. The α-AED administrated intratumorally reduced average tumor weight and increased the percentage of natural killer cells (NK), plasmatic, and plasmablast cells in mice tumors. Of note, VEGF levels in all α-AED-treated tumors was lower than in the control and vehicle groups. The tumor in situ increased response was reflected systemically by higher anti-4T1 IgG concentration in serum from α-AED-treated mice, but no other associated systemic changes were detected. The reduction in tumor size for the local injection of α-AED is associated with the anti-proliferative effect of this steroid, and the lower local levels of VEGF may be related to the imperceptible macroscopic metastasis in α-AED-treated mice. The above suggests that α-AED may be used in clinical studies to prove its efficacy as an alternative breast tumor treatment or in conjunction with already established therapies.

A new therapeutic approach for bone metastasis in colorectal cancer: intratumoral melittin.

Background: Melittin has shown antiproliferative effects on tumor cells. Therefore, it comprises a valuable compound for studies on cancer treatment. To the best of our knowledge, no studies have reported melittin effects on bone metastasis. Herein, we propose an approach based on intrametastatic melittin injection to treat bone metastases in colorectal cancer. Methods: Following the characterization of melittin and antiproliferative tests in vitro, a single dose was injected through intrametastatic route into the mouse bone metastasis model. Following treatment, metastasis growth was evaluated. Results: A single dose of melittin was able to inhibit metastasis growth. Histological analysis showed necrosis and inflammatory processes in melittin-treated metastasis. Except by mild weight loss, no other systemic effects were observed. Conclusion: Our data suggest that melittin might be a promising agent for the future development of treatment strategies aiming to reduce the bone metastasis skeletal-related impact in colorectal cancer patients with bone metastasis.

A new therapeutic approach for bone metastasis in colorectal cancer: intratumoral melittin

Abstract Background: Melittin has shown antiproliferative effects on tumor cells. Therefore, it comprises a valuable compound for studies on cancer treatment. To the best of our knowledge, no studies have reported melittin effects on bone metastasis. Herein, we propose an approach based on intrametastatic melittin injection to treat bone metastases in colorectal cancer. Methods: Following the characterization of melittin and antiproliferative tests in vitro, a single dose was injected through intrametastatic route into the mouse bone metastasis model. Following treatment, metastasis growth was evaluated. Results: A single dose of melittin was able to inhibit metastasis growth. Histological analysis showed necrosis and inflammatory processes in melittin-treated metastasis. Except by mild weight loss, no other systemic effects were observed. Conclusion: Our data suggest that melittin might be a promising agent for the future development of treatment strategies aiming to reduce the bone metastasis skeletal-related impact in colorectal cancer patients with bone metastasis.

A new therapeutic approach for bone metastasis in colorectal cancer: intratumoral melittin

Background: Melittin has shown antiproliferative effects on tumor cells. Therefore, it comprises a valuable compound for studies on cancer treatment. To the best of our knowledge, no studies have reported melittin effects on bone metastasis. Herein, we propose an approach based on intrametastatic melittin injection to treat bone metastases in colorectal cancer. Methods: Following the characterization of melittin and antiproliferative tests in vitro, a single dose was injected through intrametastatic route into the mouse bone metastasis model. Following treatment, metastasis growth was evaluated. Results: A single dose of melittin was able to inhibit metastasis growth. Histological analysis showed necrosis and inflammatory processes in melittin-treated metastasis. Except by mild weight loss, no other systemic effects were observed. Conclusion: Our data suggest that melittin might be a promising agent for the future development of treatment strategies aiming to reduce the bone metastasis skeletal-related impact in colorectal cancer patients with bone metastasis.(AU)

A Key Pathway to Cancer Resilience: The Role of Autophagy in Glioblastomas.

There are no effective strategies for the successful treatment of glioblastomas (GBM). Current therapeutic modalities effectively target bulk tumor cells but leave behind marginal GBM cells that escape from the surgical margins and radiotherapy field, exhibiting high migratory phenotype and resistance to all available anti-glioma therapies. Drug resistance is mostly driven by tumor cell plasticity: a concept associated with reactivating transcriptional programs in response to adverse and dynamic conditions from the tumor microenvironment. Autophagy, or "self-eating", pathway is an emerging target for cancer therapy and has been regarded as one of the key drivers of cell plasticity in response to energy demanding stress conditions. Many studies shed light on the importance of autophagy as an adaptive mechanism, protecting GBM cells from unfavorable conditions, while others recognize that autophagy can kill those cells by triggering a non-apoptotic cell death program, called 'autophagy cell death' (ACD). In this review, we carefully analyzed literature data and conclude that there is no clear evidence indicating the presence of ACD under pathophysiological settings in GBM disease. It seems to be exclusively induced by excessive (supra-physiological) stress signals, mostly from in vitro cell culture studies. Instead, pre-clinical and clinical data indicate that autophagy is an emblematic example of the 'dark-side' of a rescue pathway that contributes profoundly to a pro-tumoral adaptive response. From a standpoint of treating the real human disease, only combinatorial therapy targeting autophagy with cytotoxic drugs in the adjuvant setting for GBM patients, associated with the development of less toxic and more specific autophagy inhibitors, may inhibit adaptive response and enhance the sensibility of glioma cells to conventional therapies.

In Situ Gene Therapy.

Gene therapy is a technique that aims at the delivery of nucleic acids to cells, to obtain a therapeutic effect. In situ gene therapy consists of the administration of the gene product to a specific site. It possesses several advantages, such as the reduction in potential side effects, the need for a lower vector dose, and, as a consequence, reduced costs, compared to intravenous administration. Different vectors, administration routes and doses involving in situ gene transfer have been tested both in animal models and humans, with in situ gene therapy drugs already approved in the market. In this review, we present applications of in situ gene therapy for different diseases, ranging from monogenic to multifactorial diseases, focusing mainly on therapies designed for the intra-articular and intraocular compartments, as well as gene therapies for the central nervous system (CNS) and for tumors. Gene therapy finally seems to blossom as a viable therapeutic approach. The growth in the number of clinical protocols shown here is evident, and the positive outcomes observed in several clinical trials indicate that more products based on in situ gene therapy should reach the market in the next years.

La infiltración linfocitaria (tils) como factor pronóstico en pacientes con cáncer de mama Triple Negativo que realizan quimioterapia neoadyuvante / Intratumoral lymphocyte infiltration (tils) as prognostic factor in patients with Triple Negative breast cancer who underwent neoadjuvant chemotherapies

Introducción Los cánceres de mama Triple Negativo representan entre el 12 y el 17% de todos los carcinomas mamarios. Son un grupo heterogéneo con diferentes subgrupos. La reacción inflamatoria que produce el huésped como respuesta a la enfermedad puede cuantificarse a través de la infiltración linfocitaria intratumoral (tils). Objetivos Evaluar la infiltración linfocitaria intratumoral (tils) como factor pronóstico independiente en las core biopsias de las pacientes con cáncer de mama Triple Negativo que fueron sometidas a quimioterapias neoadyuvantes. Relacionarla con la respuesta patológica obtenida luego de la cirugía. Material y método Se seleccionaron retrospectivamente pacientes con carcinoma de mama Triple Negativo que realizaron quimioterapia neoadyuvante en la Unidadde Mastología de la Clínica Breast y en el Hospital Italiano de la Ciudad de La Plata entre los años 2014 y 2017. Se obtuvo una muestra de 36 pacientes. Resultados Sobre un total de 36 pacientes, 24 mostraron tils estromales menores al 50% y 12 mayores o iguales al 50%. El 16,7% de los tumores con tils menores al 50% y la mitad de los tumores con tils mayores o iguales al 50% presentaron una Respuesta Patológica Completa (rpc) post tratamiento quimioterápico neoadyuvante. En relación con los tils intratumorales, 6 tumores de 32 con tils menores al 50% (18,8%) y 4 de 4 (100%) con tils > o iguales al 50% presentaron una Respuesta Patológica Completa (rpc) post tratamiento quimioterápico neoadyuvante. Conclusiones En nuestra serie de casos, observamos que existe relación entre el porcentaje de tils y la respuesta patológica obtenida luego de la realización del tratamiento con quimioterapia neoadyuvante, siendo un factor pronóstico en las pacientes con cáncer de mama Triple Negativo

Introduction Triple Negative breast cancers account for between 12 and 17% of all breast carcinomas. They are a heterogeneous group with different subgroups. The inflammatory reaction produced by the host in response to the disease can be quantified through intratumoral lymphocyte infiltration (tils). Objectives To evaluate intratumoral lymphocyte infiltration (tils), as an independent prognostic factor, in the core biopsies of patients with Triple Negative breast cancer who underwent neoadjuvant chemotherapies. To relate it to the pathological response obtained after surgery. Materials and method Patients with Triple Negative breast carcinoma who underwent neoadjuvant chemotherapy in the Mastology Unit of the Breast Clinic and the Italian Hospital of La Plata were retrospectively selected between the years 2014 and 2017. A sample of 36 patients was obtained. Results Out of a total of 36 patients, 24 showed stromal tils less than 50% and 12 greater than or equal to 50%. 16.7% of tumours with tils less than 50% and half of tumours with tils greater than or equal to 50% presented a complete pathological response after neoadjuvant chemotherapy treatment. In relation to intratumoral tils, 6 tumors of 32 with tils less than 50% (18.8%) and 4 of 4 (100%) with tils > or equal to 50% presented a complete pathological response after neoadjuvant chemotherapy treatment. Conclusions In our series of cases, we observe that there is a relationship between the percentage of tils and the pathological response obtained after treatment with neoadjuvant chemotherapy, being a prognostic factor in patients with triple negative breast cancer

The Effects of Neoadjuvant Chemoradiation in Locally Advanced Rectal Cancer-The Impact in Intratumoral Heterogeneity.

Purpose: Intratumoral genetic heterogeneity (ITGH) is a common feature of solid tumors. However, little is known about the effect of neoadjuvant chemoradiation (nCRT) in ITGH of rectal tumors that exhibit poor response to nCRT. Here, we examined the impact of nCRT in the mutational profile and ITGH of rectal tumors and its adjacent irradiated normal mucosa in the setting of incomplete response to nCRT. Methods and Materials: To evaluate ITGH in rectal tumors, we analyzed whole-exome sequencing (WES) data from 79 tumors obtained from The Cancer Genome Atlas (TCGA). We also compared matched peripheral blood cells, irradiated normal rectal mucosa and pre and post-treatment tumor samples (PRE-T and POS-T) from one individual to examine the iatrogenic effects of nCRT. Finally, we performed WES of 7 PRE-T/POST-T matched samples to examine how nCRT affects ITGH. ITGH was assessed by quantifying subclonal mutations within individual tumors using the Mutant-Allele Tumor Heterogeneity score (MATH score). Results: Rectal tumors exhibit remarkable ITGH that is ultimately associated with disease stage (MATH score stage I/II 35.54 vs. stage III/IV 44.39, p = 0.047) and lymph node metastasis (MATH score N0 35.87 vs. N+ 45.79, p = 0.026). We also showed that nCRT does not seem to introduce detectable somatic mutations in the irradiated mucosa. Comparison of PRE-T and POST-T matched samples revealed a significant increase in ITGH in 5 out 7 patients and MATH scores were significantly higher after nCRT (median 41.7 vs. 28.8, p = 0.04). Finally, we were able to identify a subset of "enriched mutations" with significant changes in MAFs between PRE-T and POST-T samples. These "enriched mutations" were significantly more frequent in POST-T compared to PRE-T samples (92.9% vs. 7.1% p < 0.00001) and include mutations in genes associated with genetic instability and drug resistance in colorectal cancer, indicating the expansion of tumor cell subpopulations more prone to resist to nCRT. Conclusions: nCRT increases ITGH and may result in the expansion of resistant tumor cell populations in residual tumors. The risk of introducing relevant somatic mutations in the adjacent mucosa is minimal but non-responsive tumors may have potentially worse biological behavior when compared to their untreated counterparts. This was an exploratory study, and due to the limited number of samples analyzed, our results need to be validated in larger cohorts.

PTEN loss in Gleason grade 7 prostate tumors exhibits intratumoral heterogeneity and is associated with unfavorable pathological features

Background: PTEN loss is observed in 20­30% of prostate cancers and is associated with a poor outcome, but clinical details of the impact of this biomarker are unclear for intermediate grade tumors. Methods: We investigated 43 radical prostatectomy-derived grade 7 prostate tumors from the Clinics Hospital of Ribeirão Preto. Tissue microarray (TMA) blocks were constructed and PTEN copy number status was determined for all patients through fluorescence in situ hybridization (FISH). To determine the presence of PTEN protein loss in our study cohort, we performed immunohistochemistry (IHC) in TMA sections. We then developed an automated algorithm in HALO™ to identify regions of PTEN protein loss in whole prostate scanned sections from ten patients with known PTEN deletion status by FISH. Clinical analyses were conducted to determine the associations between PTEN loss and patient outcome. All statistical analyses were conducted in R v3.4.3 with P-values below 0.05 being considered statistically significant. Results: In this study of 43 grade 7 tumors, we found PTEN deletions by FISH in 18.9% of tumors, and PTEN protein loss by IHC in 16.3% of tumors. Both techniques were highly concordant and complementary. Clinical analysis demonstrated that PTEN deletion by FISH was significantly associated with positive margin invasion (P = 0.04) and Gleason score upgrade (P = 0.001). Digital image analysis of ten representative tumors demonstrated distinct intratumoral heterogeneity for PTEN protein loss in four tumors. Conclusions: This study shows that PTEN loss in Gleason grade 7 tumors can be heterogeneous and that a systematic analysis of this biomarker using a combination of FISH, IHC, and digital imaging may identify patients with a greater risk of poor outcome (AU)

O papel extracelular da proteína STI1 na biologia de glioblastomas: ênfase nos mecanismos de secreção do modelo GL261 / STI1 protein extracelular role on glioblastoma biology: enphasis on secretion mechanisms on GL-261 model

Glioblastoma é tumor primário mais frequente do sistema nervoso central e está associado a uma alta mortalidade. A formação de novos vasos (angiogênese) é um processo essencial para a manutenção desses tumores, o que torna importante a busca por terapias que interfiram neste processo, visando um melhor resultado no tratamento. A proteína stress inducible phophoprotein one (STI1) é uma co-chaperona que auxilia o dobramento de proteínas e também pode ser secretada em vesículas extracelulares (VE) e formar um complexo com a proteína príon celular (PrPC) na membrana plasmática, desencadeando diversas funções celulares, como proliferação, diferenciação e migração. O complexo STI1-PrPC também já foi associado à regulação da angiogênese em modelos de isquemia experimental, mas nada se conhece sobre o papel deste complexo na angiogênese tumoral. Um importante regulador da angiogênese é a hipóxia, que ocorre quando há diminuição dos níveis de oxigênio no tecido. Considerando a função das VEs em células tumorais e seu papel na secreção de STI1, sugere-se que a expressão e secreção de STI1 possa ser modulada pela hipóxia e contribuir na estimulação da angiogênese tumoral. Portanto esse projeto tem como objetivo investigar os mecanismos e a relevância da secreção de STI1 utilizando o modelo de glioblastoma murino GL-261. Animais transgênicos que expressam diferentes quantidades de STI1 foram inoculados com GL261 e aqueles que superexpressam a proteína (denominados TGA) apresentaram maior sobrevida e microvasculatura tumoral comprometida, com menor número de vasos totais e de menor calibre (> 20 um). A inoculação intratumoral do peptídeo STI1¬¬230-245 não alterou o crescimento tumoral de GL261 em animais tipo-selvagem e haploinsuficientes para STI1. Não houve alteração na quantidade de VE e de STI1 secretadas pro células GL261 submetidas à hipóxia. Meio condicionado e vesículas extracelulares de GL261 foram capazes de estimular a migração e formação de tubos de células endoteliais, porém ainda não foi possível determinar se a STI1 extracelular apresenta contribuição nesse fenótipo. Observamos uma tendência de pacientes com glioblastoma produzirem menor número de VE do que pacientes com astrocitomas de grau II e III, porém a diferença não foi significativa (AU)

Glioblastoma is the most frequent type of central nervous system primary tumors and are associated with a high mortality. The formation of new vessels (angiogenesis) is an essential process for the maintenance of these tumors, which makes important the search for therapies that interfere in this process, aiming at a better result in the treatment. The stress inducible phophoprotein one (STI1) is a co-chaperone that assists the folding of proteins and can also be secreted in extracellular vesicles (EV) and form a complex with the cellular prion protein (PrPC) in the plasma membrane, triggering several cellular functions such as proliferation, differentiation and migration. The STI1-PrPC complex has also been associated with the angiogenesis regulation in experimental ischemia models, but nothing is known about the role of this complex in tumor angiogenesis. An important angiogenesis regulator is the hypoxia, which occurs when tissue oxygen levels decrease. Considering the EVs role in tumor cells and their role in STI1 secretion, it is suggested that STI1 expression and secretion can be modulated by hypoxia and contribute to the stimulation of tumor angiogenesis. Therefore, this project aims to investigate the mechanisms and relevance of STI1 secretion using the GL-261 murine glioblastoma model. Transgenic animals expressing different amounts of STI1 were inoculated with GL261 and those that overexpress the protein (called TGA) presented higher survival and tumoral microvasculature with a lower number of total vessels and vessels with smaller caliber (> 20 µm). The STI1230-245 peptide intratumoral inoculation did not alter GL261 tumor growth in wild-type and haploinsufficient STI1 animals. There was no change in the amount of extracellular vesicles and STI1 secreted into GL261 cells submitted to hypoxia. Conditioned medium and extracellular vesicles of GL261 were able to stimulate the migration and formation of endothelial cell tubes, but it was not possible to determine yet if extracellular STI1 has a contribution to this phenotype. We observed a tendency for patients with glioblastoma to express less extracellular vesicles than patients with grade II and III astrocytomas, but the difference was not significant (AU)

El infiltrado tumoral del carcinoma ductal infiltrante en Holguín, Cuba / Ductal infiltrating carcinoma´s inflammatory tumor infiltrates in Holguín, Cuba

Introducción: el carcinoma ductal infiltrante es la neoplasia maligna mamaria más frecuente. Entre sus características anatomopatológicas se presenta el grado histológico de malignidad, como su factor pronóstico. Existen otras alteraciones histopatológicas, como la presencia e intensidad del infiltrado inflamatorio intratumoral y peritumoral, sobre lo cual es necesario profundizar. Objetivos: determinar la intensidad del infiltrado inflamatorio intraperitumoral y peritumoral, e identificar su relación con el grado histológico de malignidad. Métodos: el estudio de una serie de casos de 392 biopsias, diagnosticadas con carcinoma ductal infiltrante de mama, en el Departamento de Anatomía Patológica del Hospital Lenin, de Holguín, Cuba, desde el año 2011 al 2015. Se determinó la presencia e intensidad del infiltrado inflamatorio intratumoral y peritumoral, y su grado histológico de malignidad. Resultados: se logró diferenciar el 68,1% de los tumores de mama. Predominaron los infiltrados inflamatorios intratumoral y peritumoral ligeros, con 44,9% y 55,4% respectivamente. El 84,6% de los tumores bien diferenciados, mostraron un infiltrado inflamatorio intratumoral ligero. En el 47,2% de los tumores moderadamente diferenciados, y en el 41,7% de los tumores poco diferenciados, con un infiltrado inflamatorio severo; se realizaron infiltrados moderados. En los tumores bien diferenciados se observó un predominio del infiltrado inflamatorio peritumoral ligero, de hasta el 92,3%; mientras que el 48,3% de los tumores con poca diferenciación, mostraron una correlación positiva con el infiltrado inflamatorio severo. Conclusiones: en el infiltrado inflamatorio intratumoral y peritumoral, predominó la intensidad ligera, y apareció un factor pronóstico potencial: la correlación entre la intensidad del infiltrado inflamatorio y el grado histológico de malignidad.

Introduction: among most frequent malignant breast tumors, ductal carcinoma has prevailed. Its malignancy histological level has become the prognosis factor. However, histopathological abnormalities like intra and peritumoral inflammatory infiltrates intensity, still require serious studies. Objectives: to determine intra and peritumoral inflammatory infiltrates intensity, and to identify their relation to malignancy histological levels. Methods: a case series study with 392 infiltrating ductal carcinoma diagnosed biopsies, at Lenin Hospital Pathological Anatomy Department, from 2011 to 2015; in which presence and intensity of intratumoral, and peritumoral inflammatory infiltrates, and malignancy histological level, were found. Results: breast tumors were differentiated up to 68.1%. Intra and peritumoral lights inflammatory infiltrate prevailed, with 44.9% and 55.4% each. Well differentiated lights inflammatory infiltrated tumors reached 84.6%. Moderate differentiated tumors were moderate infiltrated, up to 47.2%; and poor diferentiated with severe inflamatory infíltrate, presented 41.7%. Among well differentiated tumors, peritumoral light inflamatory infiltrate revealed 92.3%. Those poor differentiated were only 48.3% positively related. Conclusions: intra and peritumoral light intensity inflammatory infiltrate was the most frequent, considered as the potential prognosis factor, because of its directly proportional relationship with malignancy histological level.

Intratumoral Bleeding in a Vestibular Schwannoma

Vestibular schwannomas (VSs) account for 70% of all tumors of the cerebellopontine angle (CPA). Their clinical presentation is often insidious, with progressive hearing loss and involvement of other cranial nerves. Spontaneous hemorrhage in those tumors is very unusual, and generally presents with acute clinical features such as nausea, vomiting, headache and altered consciousness, usually with marked dysfunction of the cranial nerve involved, and with new deficits of neighboring cranial nerves. Asymptomatic patients are extremely rare. We present a case report of an incidental VS with asymptomatic bleeding, which evolved to death after surgery.

Schwannomas vestibulares (SVs) são responsáveis por cerca de 70% de todos os tumores do ângulo pontocerebelar. Sua apresentação costuma ser insidiosa, com perda auditiva progressiva e envolvimento de outros nervos cranianos. Hemorragia espontânea nesses tumores é incomum, e geralmente apresenta-se agudamente, com náusea, vômitos, cefaleia e alterações de consciência, normalmente com disfunção importante dos nervos cranianos envolvidos e com novos déficits dos nervos próximos. Pacientes assintomáticos são extremamente raros. Apresentamos um relato de caso de um SV incidental com sangramento assintomático que evoluiu para o óbito após cirurgia.

Intratumoral heterogeneity of CD44v6 in rectal cancer.

PURPOSE: CD44v6 plays a controversial role in tumor progression and patient outcome in colorectal cancer by plenty of conflicting reports. The purpose of this study was to profile the intratumoral heterogeneity of CD44v6 in rectal cancer and investigate its role in lymph node metastasis. METHODS: Sixty patients were included in this study. Immunohistochemistry for CD44v6 was performed in normal mucosa, primary tumor, and lymph node metastasis with whole tissue sections. The staining intensity in tumor center and invasive front was separately measured. Sampling bias was evaluated by quantitative real-time PCR with 15 pairs of frozen tissues from different sites of the primary tumor. RESULTS: CD44v6 expression increased from normal mucosa to primary tumor to lymph node metastasis. Multiple intratumoral staining patterns was observed in primary tumor, and CD44v6 expression in invasive front was significantly higher than that in tumor center. In addition, mRNA expression levels differed across different geographical regions of the tumor. No association between CD44v6 expression and lymph node metastasis was revealed. CONCLUSIONS: Substantial intratumoral heterogeneity of CD44v6 exists in rectal cancer that impacts the outcome of individual studies. CD44v6 expression should be assessed in a more precise way with a specified staining pattern and in a designated location.

Comparative prognostic relevance of breast intra-tumoral microvessel density evaluated by CD105 and CD146: A pilot study of 42 cases.

UNLABELLED: Angiogenesis is a key process for metastatic progression. While it has been established that the evaluation of breast tumoral microvessel density by CD105 marker is a potential prognostic parameter, its evaluation by CD146 marker has been poorly studied. AIM: The purpose of this study was to compare the prognostic value of intra-tumoral microvessel density assayed by CD105 and CD146 in early breast cancer patients. METHODS: 42 women with breast infiltrative ductal carcinoma (I and II-stages) were retrospectively reviewed. Intra-tumoral microvessel density was immunohistochemically examined using antibodies anti-CD105 and CD146 in paraffin-embedded tissues, and their association with classical prognostic-markers, metastatic recurrence, metastasis-free survival and overall survival was analyzed. RESULTS: High microvessel density assessed by CD146 was significantly associated with a higher risk of developing metastasis (p=0.0310) and a shorter metastasis-free survival (p=0.0197). In contrast, when we used the CD105-antibody, we did not find any significant association. Finally, CD146 showed to be an independent predictive indicator for metastasis-free survival (p=0.0055). CONCLUSION: Our data suggest that the intra-tumoral microvessel density evaluated by CD146 may be a more suitable predictor of metastatic development than that evaluated by CD105 in early breast cancer.

In vivo assessment of intratumoral aspirin injection to treat hepatic tumors.

AIM: To study the antineoplastic efficacy of 10% aspirin intralesional injection on VX2 hepatic tumors in a rabbit model. METHODS: Thirty-two male rabbits (age: 6-9 wk; body weight: 1700-2500 g) were inoculated with VX2 hepatic tumor cells (10(4) cells/rabbit) via supra-umbilical median laparotomy. On day 4 post-implantation, when the tumors were about 1 cm in diameter, the rabbits were randomly divided into the following groups (n = 8 each group) to assess early (24 h) and late (7 d) antineoplastic effects of intratumoral injection of 10% bicarbonate aspirin solution (experimental groups) in comparison to intratumoral injection of physiological saline solution (control groups): group 1, 24 h control; group 2, 24 h experimental; group 3, 7 d control; group 4, 7 d experimental. The serum biochemistry profile (measurements of glycemia, alkaline phosphatase, gamma-glutamyl transferase, aspartate aminotransferase, and alanine aminotransferase) and body weight measurements were obtained for all animals at the following time points: D0, before tumor implant; D4, day of treatment; D5, day of sacrifice for groups 1 and 2; D11, day of sacrifice for groups 3 and 4. Gross assessments of the abdominal and thoracic cavities were carried out upon sacrifice. The resected liver tissues, including hepatic tumors, were qualitatively (general morphology, signs of necrosis) and quantitatively (tumor area) assessed by histopathological analysis. RESULTS: Gross examination showed no alterations, besides the left hepatic lobe tumors, had occurred in the thoracic and abdominal cavities of any animal at any time point evaluated. However, the features of the tumor foci were distinctive between the groups. Compared to the control groups, which showed normal unabated tumor progression, the aspirin-treated groups showed imprecise but limited tumor boundaries and a general red-white coloration (indicating hemorrhaging) at 24 h post-treatment, and development of yellow-white areas of a cicatricial aspect at 7 d after treatment. At all time points evaluated, all except one biochemical parameters tested within the reference range (P > 0.05); a significant increase was detected in the alkaline phosphatase level of the control group 3 on D11 (P < 0.05). At 24 h post-treatment, the aspirin-treated groups showed extensive coagulation necrosis accompanied by a remarkable absence of viable tumor foci; at 7 d after treatment, the tumors had completely disappeared in these animals and fibrous necrotic nodules had developed. In contrast, throughout the study course, the tumors of the control groups remained unchanged, showing tumor nodules without necrosis at the time point corresponding to 24 h post-treatment and increased amounts of tumor nodules at the time point corresponding to 7 d post-treatment. Quantitative analysis of the remaining tumor area revealed that the aspirin-treated groups had significantly smaller tumor foci at 24 h post-treatment (8.5% ± 0.7%) and at 7 d after treatment (11.0% ± 4.2%), compared to those in the control groups (24 h: 98.5% ± 1.5% and 7 d: 94.0% ± 2.7%; both, P < 0.005). CONCLUSION: Intralesional injection of a 10% aspirin solution causes destruction of VX2 hepatic tumors in rabbits without evidence of relapse at 7 d after treatment administration.

Cystic craniopharyngioma: intratumoral chemotherapy with alpha interferon / Craniofaringioma cístico: quimioterapia intratumoral com interferon alfa

OBJECTIVE: To assess whether the cystic craniopharyngiomas can be controlled with the use of intratumoral applications of interferon alpha. METHOD: Nineteen patients with the diagnosis of cystic craniopharyngioma were treated with intratumoral chemotherapy with interferon alpha from January 2002 to April 2006. All patients underwent placement of an intracystic catheter connected to an Ommaya reservoir. Through this reservoir were made applications during chemotherapy cycles. Each cycle corresponded to application of 3,000,000 units of interferon alpha three times per week on alternate days totalizing 36,000,000 units. Response to treatment was evaluated by calculating the tumor volume on MRI control after one, three and six months after the end of each cycle. Patients who developed worsening of symptoms or who had insignificant reduction in tumor volume during follow-up underwent repeat cycle chemotherapy. RESULTS: Four patients received four cycles of chemotherapy, three patients received three cycles, six patients received two cycles and six patients received one. The lower percentage of reduction in tumor volume was 60 percent and the bigger reduction was 98.37 percent. Eleven patients had a reduction greater than 90 percent. Five patients had a tumor reduction between 75 and 90 percent and in three patients the tumors were reduced by less than 75 percent. No deaths occurred during treatment and side effects of interferon alpha were well tolerated. No treatment was discontinued. Follow-up after the last application ranged from one year and five months to three years and nine months. CONCLUSION: The intratumoral chemotherapy with interferon alpha decreases the volume of cystic craniopharyngiomas and so far can be considered a new therapeutic alternative.

OBJETIVO: Avaliar se os craniofaringiomas císticos podem ser controlados com aplicações intratumorais de interferon alfa. MÉTODO: De janeiro de 2002 a abril de 2006, 19 pacientes foram submetidos à colocação de um cateter intracístico conectado a reservatório de Ommaya para aplicações intratumorais de ciclos de 36.000.000 de unidades de interferon alfa. A resposta ao tratamento foi avaliada pelo cálculo do volume tumoral na ressonância magnética de controle ao término de cada ciclo. RESULTADOS: Os pacientes receberam de um a quatro ciclos de quimioterapia. Onze pacientes apresentaram uma redução do volume tumoral maior que 90 por cento; cinco pacientes apresentaram uma redução entre 75 por cento e 90 por cento e três pacientes uma redução menor de 75 por cento. Não houve óbitos durante o tratamento e os efeitos colaterais do inferferon alfa foram bem tolerados. Nenhum tratamento foi interrompido. CONCLUSÃO: A quimioterapia intratumoral com interferon alfa diminui o volume dos craniofaringeomas císticos e pode ser considerada uma nova alternativa terapêutica.

Prognostic value of podoplanin expression in intratumoral stroma and neoplastic cells of uterine cervical carcinomas

OBJECTIVE: To investigate the clinicopathological significance of podoplanin expression in the intratumoral stroma and neoplastic cells of early stage uterine cervical cancer. MATERIALS AND METHODS: A total of 143 patients with clinical stage I and IIA uterine cervical carcinomas underwent surgery between 2000 and 2007. Clinicopathological data and slides associated with these cases were retrospectively reviewed. Immunodetection of podoplanin expression in histologic sections of tissue microarray blocks was performed using the monoclonal antibody D2-40. RESULTS: Expression of podoplanin was detected in neoplastic cells in 31/143 (21.6 percent) cases, with 29/31 (93.5 percent) of these cases diagnosed as squamous carcinoma. For all of the cases examined, the strongest signal for podoplanin expression was observed at the proliferating edge of the tumor nests. The rate of positive podoplanin expression for node-positive cases was lower than that of node-negative (18.9 percent vs. 22.6 percent, respectively). Furthermore, the rate of positive podoplanin expression in fatal cases was 10.5 percent vs. 21.6 percent, respectively. In 27/143 (18.8 percent) cases, podoplanin expression was detected in fibroblasts of the intratumoral stroma, and this expression did not correlate with patient age, clinical stage, tumor size, histologic type, depth of infiltration, or vascular involvement. Moreover, expression of podoplanin in intratumoral stroma fibroblasts was only negatively associated with nodal metastasis. A greater number of fatal cases was observed among negative intratumoral stroma fibroblasts (15.5 percent vs. 3.7 percent, respectively), although this difference was not significant. CONCLUSIONS: These preliminary results suggest that podoplanin may have a role in host-tumor interactions and, as a result, may represent a favorable prognostic factor for squamous cervical carcinomas.