Updated guidelines for predictive biomarker testing in advanced non-small-cell lung cancer: a National Consensus of the Spanish Society of Pathology and the Spanish Society of Medical Oncology.

In 2011 the Spanish Society of Medical Oncology (SEOM) and the Spanish Society of Pathology (SEAP) started a joint project to establish guidelines on biomarker testing in patients with advanced non-small-cell lung cancer (NSCLC) based on current evidence. As this field is constantly evolving, these guidelines have been updated, previously in 2012 and 2015 and now in 2019. Current evidence suggests that the mandatory tests to conduct in all patients with advanced NSCLC are for EGFR and BRAF mutations, ALK and ROS1 rearrangements and PD-L1 expression. The growing need to study other emerging biomarkers has promoted the routine use of massive sequencing (next-generation sequencing, NGS). The coordination of every professional involved and the prioritisation of the most suitable tests and technologies for each case remains a challenge.

Relationship Between Albuminuria During the First Year and Antibody-Mediated Rejection in Protocol Biopsies in Kidney Transplant Recipients.

BACKGROUND: Antibody-mediated rejection is the main cause of deterioration of kidney transplants and frequently is detected only by means of protocol biopsies. The aim of this study was to relate the presence of albuminuria throughout the 1st year to the histologic findings detected by 1-year protocol biopsies in kidney graft recipients. METHODS: Retrospective observational study of 86 protocol biopsies 1 year after transplantation. Albuminuria was measured at 3, 6, 9, and 12 months in urine samples and expressed as albumin/creatinine (mg/g). RESULTS: Analysis of biopsies, reflected according to the Banff criteria, the following categories: fibrosis and tubular atrophy, 35 (40.7%); cellular rejection, 13 (15.1%); antibody-mediated rejection, 8 (9.3%); chronic glomerulopathy, 10 (11.6%); normal, 14 (16.3%); recurrence, 1 (1.2%); and other, 5 (5.8%). The proportions of patients with albuminuria for Banff scale scores (0 vs ≥1, respectively) at 6 and 12 months, respectively, after transplantation, were: for marker glomerulitis, 45.5% versus 59.3% (P = .021) and 36.4% versus 70.4% (P < .001); for marker glomerulopathy, 49.1% versus 50.0% (P = .051) and 42.1% versus 58.3% (P = .019); for marker peritubular capillaritis, 45.8% versus 60.9% (P = .047) and 39.0% versus 69.6% (P = .276); and for marker C4d, 49.2% versus 56.3% (P = .894) and 46.2% versus 56.3% (P = .774). CONCLUSIONS: The presence of albuminuria after renal transplantation is common, especially in patients with proteinuria. Persistent albuminuria after transplantation, even at low levels, can be indicative of subclinical antibody-mediated rejection. Additional broader studies to relate the albuminuria to histologic changes observed in protocol biopsies are required.

Behaviour and survival of high-grade neuroendocrine carcinomas of the lung.

INTRODUCTION: Large-cell neuroendocrine carcinoma is an aggressive variant of large-cell carcinoma of the lung, which has poor survival in most series, resembling that of small-cell lung carcinoma. We report our retrospective assessment of surgically-resected cases of both tumours. METHODS: 33 large-cell neuroendocrine carcinomas and 16 peripheral small-cell lung carcinomas were reassessed retrospectively. Survival rates of both tumours in surgically-resected cases were calculated and compared using Kaplan-Meier survival curves and Log Rank test, respectively. RESULTS: In large-cell neuroendocrine carcinomas, there were 25 patients with pathologic stage I, 4 with pathologic stage II and 4 with pathologic stage III. In small-cell lung carcinomas, there were 6 patients with pathologic stage I, 3 with pathologic stage II and 7 with pathologic stage III. 12% of large-cell neuroendocrine carcinomas and 62.5% of small-cell lung carcinomas were of advanced disease. The mean follow-up was 89 months. The actuarial survival for the 2 groups was not significantly different. CONCLUSION: Large-cell neuroendocrine carcinomas of the lung have poor prognosis even in early stages, with survival rates similar to that of small-cell lung carcinomas.

[Hormone expression and opioid receptors in fetal and adult lung]. / Expresión hormonal y de receptores opioides en pulmón fetal y del adulto.

OBJECTIVES: To describe the cellular distribution and level of expression of certain hormones and opioid receptors during fetal development and in the lung of the healthy adult. METHOD: We sampled lung tissue from fetuses at three stages of development (pseudoglandular, canalicular and saccular) (3 samples per stage), from newborn infants (3), from 10-month-old infants (2) and from adults (3) who had died without lung disease. After specific immunohistochemical staining for hormones (calcitonin, parathormone, serotonin and adrenocorticotropic hormone - ACTH) and opioid receptors, we assessed the percentage of positive cells for each cell type in each sample. RESULTS: Serotonin is the first to appear (pseudoglandular stage in isolated neuroendocrine cells) and it disappears later. Calcitonin appears in the canalicular stage in neuroendocrine and lung cells. Expression is at its peak at birth and is less in the adult lung. We found no ACTH or parathormone production. Opioid receptors appear in the canalicular stage and peak at birth. In adult lung, bronchiolar muscle and mesothelial cells, only delta-type opioid receptors are present. CONCLUSIONS: Pulmonary hormone secretion is significant during fetal development and peaks at birth. Calcitonin is the main hormone produced in the fetal lung. Opioid receptors are present during fetal development in various types of cells and peak at birth. An understanding of the expression of active substances could have therapeutic relevance in certain conditions, such as bronchial asthma or respiratory distress syndrome in the child.

Expresión hormonal y de receptores opioides en pulmón fetal y del adulto / Hormone expression and opioid receptors in fetal and adult lung

OBJETIVOS: Describir la distribución celular y el grado de expresión de diversas hormonas y receptores de opioides en el desarrollo embrionario y en el pulmón sano del adulto. MÉTODO: Seleccionamos tejido pulmonar de las tres etapas del desarrollo fetal (seudoglandular, canalicular y sacular, tres muestras por etapa), de recién nacidos (tres), niños de 10 meses (dos) y adultos (tres) fallecidos sin afección pulmonar. Practicamos tinción inmunohistoquímica para hormonas específicas (calcitonina, parathormona, serotonina y hormona adrenocorticotropa [ACTH]) y receptores de opioides tipo delta y mu. Valoramos el porcentaje de células positivas así como el tipo celular reactivo en cada caso. RESULTADOS: La serotonina es la primera en aparecer (estadio seudoglandular en células neuroendocrinas aisladas) para posteriormente desaparecer. La calcitonina aparece en el estadio canalicular en células neuroendocrinas y neumocitos. Su expresión máxima es al nacimiento y disminuye en el pulmón adulto. No hemos encontrado producción de ACTH ni de parathormona. Los receptores de opioides aparecen en la fase canalicular y alcanzan el máximo grado en el nacimiento. En el adulto sólo existen receptores para opioides tipo delta en neumocitos, células musculares bronquiolares y mesoteliales. CONCLUSIONES: La hormonosecreción pulmonar es importante durante el desarrollo fetal y alcanza su máxima expresión en el nacimiento. La principal hormona que produce el pulmón fetal es la calcitonina. Existen receptores opioides durante el desarrollo fetal en diferentes tipos celulares y alcanzan su máxima expresión al nacimiento. El conocimiento de la expresión de sustancias activas podría tener consecuencias terapéuticas en determinados procesos patológicos como el síndrome de distrés respiratorio en el niño o el asma bronquial (AU)

Prichard's structures of the fossa ovalis are not histogenetically related to cardiac myxoma.

AIMS: Cardiac myxomas are neoplasms of unknown histogenesis. They are thought to arise from hypothetical subendothelial vasoformative reserve cells or from primitive cells which reside in the fossa ovalis and surrounding endocardium. In 1951 Prichard described a kind of microscopic endocardial structure with a predilection for the interatrial septum, which were suggested to be related to cardiac myxomas. To confirm the existence of Prichard's structures and to clarify their role in the genesis of cardiac myxomas, we examined histologically the fossa ovalis and we performed an immunohistochemical study of the endocardial abnormalities that were found. METHODS AND RESULTS: A prospective histological study of 100 interatrial septa and an immunohistochemical study of three out of the 12 endocardial abnormalities that were detected, as well as of four conventional cardiac myxomas were accomplished. Antibodies were used to vimentin, CD31, CD34, alpha-smooth muscle actin, S100 protein, thrombomodulin, calretinin and c-kit (CD117), a tyrosine kinase growth factor receptor for stem cell factor usually expressed by embryonic/fetal endothelium. Structures similar to the ones described by Prichard were found in 12% of septa, most of them in the left side of the fossa ovalis. The hearts with these structures were from patients 10 years older than the ones without them (72 +/- 10 versus 62 +/- 16 years, P=0.006). Immunohistochemically the cells comprising Prichard's structures were positive for vimentin, CD31, CD34 and thrombomodulin, and negative for alpha-smooth muscle actin, S100 protein, calretinin and c-kit. Therefore these cells seem to be mature endothelial cells, but not primitive multipotential mesenchymal cells. Furthermore, these cells were not found in the atrial tissue from the bases of any of the conventional cardiac myxomas. CONCLUSIONS: Our study suggests that there is no apparent relation between Prichard's structures and cardiac myxomas, and that Prichard's minute endocardial deformities are age-related phenomena.

Thrombomodulin, calretinin and c-kit (CD117) expression in cardiac myxoma.

The immunohistochemical profile of cardiac myxoma has been debated. The tumor is thought to be derived from multipotential undifferentiated mesenchymal cells. A consistent marker for this tumor has not been found. In this article an immunohistochemical study of 23 cardiac myxomas was accomplished. This study comprised the immunoreactivity of the tumors for thrombomodulin, calretinin and and c-kit (CD117). To the best of our knowledge, thrombomodulin and c-kit have not been tested in cardiac myxoma. Calretinin expression has been recently demonstrated in cardiac myxoma, although this finding has not been yet validated. Surface lining cells, tumor vascular endothelium, cells around the vascular slits and stromal cells embedded in the myxoid matrix were assessed independently. All tumors showed reactivity for thrombomodulin in the surface cells and in the endothelium of neoplastic vessels. 82.6% of cardiac myxomas expressed thrombomodulin in the stromal cells and 69.6% of the tumors were reactive in the perivascular cells. 73.9% of cardiac myxomas expressed calretinin in the stromal cells and in the perivascular cells. All myxomas were negative for c-kit. Thrombomodulin and calretinin may be important diagnostic aids for cardiac myxoma. Cardiac myxoma cells do not express embryonic/fetal endothelial antigens.

Human herpesvirus-8 genes are expressed in pulmonary inflammatory myofibroblastic tumor (inflammatory pseudotumor).

The presence of human herpesvirus-8 DNA sequences, as well as an overexpression of human interleukin-6 and human cyclin D1 in myofibroblastic cells of inflammatory myofibroblastic tumor (inflammatory pseudotumor), has recently been reported. We describe the pattern of human herpesvirus-8 gene expression in five cases of pulmonary inflammatory myofibroblastic tumor. Reverse transcriptase-polymerase chain reaction (RT-PCR), with several positive and negative controls, was performed to detect mRNA of 11 open reading frames encoded by human herpesvirus-8 in lytic and latent stages of viral replicative cycle. We found molecular transcripts from ORF16, ORFK13, and ORF72 in the five cases and from ORFK2 in four of five neoplasms. The corresponding encoded proteins were human homologous oncoproteins (viral cyclin-D), inflammatory cytokines (viral IL-6), and inhibitors of apoptotic pathways (viral FLIP and viral Bcl-2), mostly expressed in a latent viral replicative stage. The rest of open reading frames examined included mainly lytic-associated genes and showed no expression. The spectrum of expressed viral genes is not the same as can be observed in Kaposi's sarcoma or multicentric Castleman's disease, suggesting that human herpesvirus-8 plays a different role in the pathogenesis of its associated diseases. These differences may be related to either cell-specific or immunologic host factors.

Papillary adenoma of the kidney with mucinous secretion.

Although infrequently, mucin secretion has previously been reported in papillary renal cell carcinoma. We here investigate the presence of mucin in a series of 93 renal papillary adenomas in 58 patients. Acid mucin was present in four cases (4.3% of the tumors; 6.9% of the patients), in which basophilic mucin secretion was evident with hematoxylin-eosin. To the best of our knowledge mucin secretion has not been reported in renal papillary adenoma. We describe two different types of mucin secretion: intracytoplasmic and luminal. The secretion was intracellular in numerous scattered tumor cells in two cases, focal luminal in one case, and mixed intracellular and luminal in another case. Mucin production, despite its low frequency, can be considered as an additional feature of renal papillary adenoma. Mucin production suggests that renal papillary adenoma and papillary renal cell carcinoma are actually not two independent biological processes, but a continuum of one biological process.

Simultaneous multiorgan presence of human herpesvirus 8 and restricted lymphotropism of Epstein-Barr virus DNA sequences in a human immunodeficiency virus-negative immunodeficient infant.

Because a profound dysregulation of the immune system occurs in primary immunodeficiencies, viral infections are not uncommon. Human herpesvirus (HHV)-8 DNA was detected by polymerase chain reaction (PCR) analysis, Southern blotting, and in situ hybridization (ISH) in peripheral blood mononuclear cells and lymphoid organs (bone marrow, spleen, and lymph nodes) and endothelial and epithelial cells and macrophages from several organs (skin, lung, esophagus, intestine, choroid plexus [but not in brain or cerebellum], heart, striated muscle, liver, and kidney) of a human immunodeficiency virus-negative infant with DiGeorge anomaly who died of disseminated infection. Epstein-Barr virus DNA sequences were detected in the spleen and lymph nodes (by PCR and ISH) and in bone marrow (only by ISH) but not in blood or nonlymphoid organs. This report is believed to be the first of multiorgan dissemination of HHV-8 in a primary immunodeficiency.

Presence of human herpesvirus-8 DNA sequences and overexpression of human IL-6 and cyclin D1 in inflammatory myofibroblastic tumor (inflammatory pseudotumor).

Inflammatory myofibroblastic tumor (IMT) is composed of myofibroblasts, plasma cells, and lymphocytes. Cytokines are possibly involved in its pathogenesis. Human herpesvirus-8 (HHV-8) encodes cell cycle regulatory and signaling proteins. A combination of nested PCR with several negative controls and Southern blot methods showed the presence of HHV-8 DNA in seven cases of IMT. Additionally, strong expression was demonstrated by in situ hybridization in many tumoral nuclei. Most of the myofibroblasts in all of the cases were immunoreactive for human IL-6 and cyclin D1. These cytokines probably have a paracrine action and may sustain myofibroblastic growth. HHV-8 could play an essential role in triggering IMT development by a local reactivation of viral lytic replication. The relationship between HHV-8 and immunosuppression status as the only associated cause for tumorigenesis should be revised.

Telomerase activity in pulmonary neuroendocrine tumors: correlation with histologic subtype (MS-0060).

The authors measured telomerase activity using the telomeric repeat amplification protocol-enzyme-linked immunosorbent assay method in 13 neuroendocrine pulmonary neoplasms and in non-neoplastic frozen lung samples from the same patients. These cases belonged to the complete neuroendocrine neoplastic spectrum: four typical carcinoids, three atypical carcinoids, four large cell neuroendocrine lung carcinomas, and two small cell lung carcinomas. The authors performed the same assay for 52 non-neoplastic lung tissues from the surgical files in their department (negative controls). They verified the presence (or absence) of neoplastic tissue in every case by looking at one frozen section done in the same tissue used for telomerase assay. The telomerase activity level in non-neoplastic tissues (mean, 182 A450nm U) was similar to that obtained in the typical carcinoids (mean, 104.5 A450nm U). All neuroendocrine tumors but the typical carcinoids showed high levels of telomerase activity (mean, 1,750.8 A450nm U). According to the telomerase hypothesis, typical carcinoid cells are mortal pre-M2 stage cells, but atypical carcinoid, large cell neuroendocrine lung carcinoma, and small cell lung carcinoma cells are immortal post-M2 stage cells. This finding may be of important prognostic significance in these kinds of tumors. Measurement of enzyme activity with a good morphologic control could be necessary in telomerase activity assay.

Spindle cell rhabdomyosarcoma in adults. A case report and literature review.

Spindle cell rhabdomyosarcoma, a recently described variant of embryonal rhabdomyosarcoma in children, carries a favorable prognosis when compared with other types of rhabdomyosarcoma. This tumor is rare in adults, and only four cases have been documented previously. The clinicopathological study of such a case occurring in the retroperitoneal space of a 53-year-old man is herein reported. The patient died of uncontrolled local recurrence and hepatic metastases seven months after diagnosis. Based on the analysis of the data of the five cases reported, including the present one, it can be stated that spindle cell rhabdomyosarcoma in adults is not associated with the favorable outcome observed in the pediatric population.

Presence of human herpesvirus 8 DNA sequences in renal transplantation-associated pleural Kaposi sarcoma.

OBJECTIVE: To describe one case of symptomatic skin and pleural Kaposi sarcoma (KS) associated with kidney transplantation. Diagnosis was supported by morphologic study and human herpesvirus 8 (HHV-8) detection in both tissues. Pulmonary involvement was not present. DESIGN: The presence of HHV-8 DNA sequences was proved using polymerase chain reaction (PCR), Southern blot hybridization, and in situ hybridization. SETTING: Human herpesvirus 8 is found in most KS from patients with and without the acquired immunodeficiency syndrome. Clinically significant pulmonary infiltration by KS is diagnosed uncommonly antemortem, and pleural disease is exceptional. PATIENT: A 49-year-old man who had renal transplant with immunosuppressive therapy (tacrolimus and prednisone) and developed a cutaneous KS. A pleural effusion appeared without pulmonary involvement. Both lesions disappeared when immunosuppressive drugs were suspended. Later, the pleural effusion and the cutaneous lesions reappeared. Pleural biopsy specimens showed KS infiltration. OUTCOME: The patient refused treatment and was lost to follow-up. RESULTS: The skin and pleural biopsies showed a proliferation of spindle-shaped cells positive for CD34. The HHV-8 sequences were detected by nested PCR. No amplification was detected in uninvolved skin from the patient or in peripheral blood mononuclear cells from 10 healthy individuals used as controls. The Southern blot hybridization confirmed these results. CONCLUSIONS: To our knowledge, this is the first report of HHV-8 in symptomatic pleural KS, which was probably associated with immunosuppression after kidney transplantation. The demonstration of HHV-8 DNA in biopsy material in the appropriate cells could be diagnostic when the morphologic setting is consistent with KS.

Papillary (chromophil) renal cell carcinoma with mucinous secretion.

We investigated the presence of mucin in a series of 20 papillary renal cell carcinomas. Acid mucin was present in three cases (15%), in which basophilic mucin secretion was evident with hematoxylin-eosin. This secretion reacted positively with Mayer's mucicarmine, Alcian blue and high-iron diamine, but was negative for PAS in all the cases, indicating the presence of sulphomucins. We describe two different types of mucin secretion: luminal and intracytoplasmic. The secretion was abundant, diffuse or extensive, luminal in two cases and intracellular in numerous scattered tumor cells in one case. All tumors were < 3 cm in diameter (low grade, stage I). In the three mucin-secreting papillary tumors mentioned above, the immunohistochemical and lectin studies indicate both a proximal and a distal tubular staining pattern. Mucinous secretion in these tumors can be ascribed either to modulation or direct metaplasia of the tumor epithelium. Mucin production, despite its low frequency, can be considered an additional feature of papillary renal cell carcinoma. Thus, the presence of luminal or intracytoplasmic mucin deposition does not exclude papillary renal cell carcinoma from the differential diagnosis in cases of intra- or extrarenal carcinomas.

Primary lymphoplasmacytoid lymphoma of the trachea with immunoglobulin G paraprotein.

A primary tracheal lymphoma with immunoglobulin G (IgG)-associated monoclonal serum paraprotein treated with surgery and chemotherapy is reported. As far as we know this is the first lymphoplasmacytoid lymphoma reported in the tracheobronchial tree and the first with a serum and tissue IgG monoclonal paraprotein. Differential diagnosis must be made essentially with extramedullary plasmacytoma and mucosa-associated lymphoid tissue lymphoma. CD-45RB strong positivity and the absence of lymphoepithelial lesions may help to differentiate lymphoplasmacytoid lymphoma from them. We expand the spectrum of lymphoid lesions with plasmacytoid features that can occur in the tracheobronchial tract.

Sarcomatoid chromophobe cell renal carcinoma: immunohistochemical and lectin study in one case.

We present one case of sarcomatoid chromophobe cell renal carcinoma with an indolent clinical course and assume that the carcinomatous component may affect the biologic behavior. The patient was a 61-year-old man who underwent right radical nephrectomy for a 11.2 cm tumor in the lower pole. The immunohistochemical findings demonstrate that EMA and cytokeratins 8 and 18 are useful markers for the sarcomatoid fraction, and the lectin study shows a loss of surface blood antigen. Chromophobe cell carcinoma may convert into sarcomatoid carcinoma. The existence of sarcomatoid renal cell carcinoma as a distinct entity should be re-considered.

Chromophobe cell renal carcinoma: DNA flow cytometry and proliferation status in 9 cases.

Nine cases of chromophobe cell renal carcinoma are reported, 2 in pregnant women. One neoplasm progressed and the patient died of disease, and this tumor presented the highest mitotic index and focal positivity to Ki-67 antibody. DNA flow-cytometric study discovered 89% of aneuploidy, with 3 tetraploid cases. We conclude that chromophobe cell renal carcinoma is a relatively indolent but clearly malignant neoplasm. We suggest that the mitotic index could be the best method to predict malignant behavior. The high incidence of aneuploid cell lines may be due to structural anomalies; it is not indicative of malignant behavior.