ABSTRACT
BACKGROUND: There is a strong unmet need to improve systemic therapy in mesothelioma. Chemotherapy with cisplatin and pemetrexed improves survival in malignant pleural mesothelioma, and immune checkpoint inhibitors are an emerging treatment in this disease. We aimed to evaluate the activity of durvalumab, an anti-PD-L1 antibody, given during and after first-line chemotherapy with cisplatin and pemetrexed in patients with advanced malignant pleural mesothelioma. METHODS: DREAM was a multicentre, single-arm, open-label, phase 2 trial done in nine hospitals in Australia. Eligible patients were aged 18 years or older and had histologically confirmed malignant pleural mesothelioma considered unsuitable for cancer-directed surgery, an Eastern Cooperative Oncology Group performance status of 0 or 1, and measurable disease as per the modified Response Evaluation Criteria in Solid Tumors version 1.0 (mRECIST) for mesothelioma that was previously untreated with systemic therapy. All histological subtypes were eligible. The first six participants were treated for two cycles in a safety run-in. All participants received cisplatin 75 mg/m2, pemetrexed 500 mg/m2, and durvalumab 1125 mg intravenously on day 1 of a 3-weekly schedule for a maximum of six cycles. Change from cisplatin to carboplatin with an area under the curve of 5 was permitted. Durvalumab was continued for a maximum of 12 months. The primary endpoint was progression-free survival at 6 months, measured according to mRECIST for malignant pleural mesothelioma and analysed in the intention-to-treat population. Safety analyses included all participants who receive at least one dose of any study drug. This study is registered with the Australia New Zealand Clinical Trials Registry, ACTRN12616001170415. FINDINGS: Between Dec 28, 2016, and Sept 27, 2017, 55 participants were enrolled. 54 patients were eligible and were followed up for a median of 28·2 months (IQR 26·5-30·2). 31 (57%; 95% CI 44-70) of 54 patients were alive and progression-free at 6 months. The most common grade 3-4 adverse events were neutropenia (seven [13%] patients), nausea (six [11%]), and anaemia (four [7%]). A total of 60 serious adverse events occurred in 29 participants, five of which were considered possibly related to durvalumab. Five patients died during the study treatment; none of these five deaths were attributed to study treatment. INTERPRETATION: The combination of durvalumab, cisplatin, and pemetrexed has promising activity and an acceptable safety profile that warrants further investigation in a randomised phase 3 trial. FUNDING: AstraZeneca.
Subject(s)
Antibodies, Monoclonal/administration & dosage , Cisplatin/administration & dosage , Lung Neoplasms/drug therapy , Mesothelioma/drug therapy , Pemetrexed/administration & dosage , Pleural Neoplasms/drug therapy , Adolescent , Adult , Aged , Antibodies, Anti-Idiotypic/administration & dosage , Antibodies, Anti-Idiotypic/adverse effects , Antibodies, Monoclonal/adverse effects , Antineoplastic Combined Chemotherapy Protocols/administration & dosage , Antineoplastic Combined Chemotherapy Protocols/adverse effects , Australia/epidemiology , B7-H1 Antigen/antagonists & inhibitors , B7-H1 Antigen/genetics , Cisplatin/adverse effects , Female , Humans , Lung Neoplasms/genetics , Lung Neoplasms/immunology , Lung Neoplasms/pathology , Male , Mesothelioma/genetics , Mesothelioma/immunology , Mesothelioma/pathology , Mesothelioma, Malignant , Middle Aged , Pemetrexed/adverse effects , Pleural Neoplasms/genetics , Pleural Neoplasms/immunology , Pleural Neoplasms/pathology , Progression-Free SurvivalABSTRACT
Immune checkpoint inhibition is an important therapeutic option in patients with non-small cell lung cancer. Programmed cell death ligand-1 (PD-L1) expression may serve as a predictive marker for anti-PD-1/PD-L1 therapies. The relationship between non-small cell lung cancer PD-L1 expression and clinicopathological characteristics remains unclear and there is no population level Australian data. We report the results of PD-L1 testing in patients with non-small cell lung cancer diagnosed at major Western Australian public hospitals served by a single state Pathology provider. We analyzed PD-L1 expression by immunohistochemistry in 241 non-small cell lung cancer specimens using the 22C3 clone on a Dako autostainer platform. Tumor cell PD-L1 expression was scored as Tumor Proportion Score and categorized using pre-specified subsets of 1%, 1-49% and ≥ 50% for correlation with clinicopathologic features. PD-L1 Tumor Proportion Score was 1% in 65 (27%) cases, 1-49% in 100 (41%) cases and ≥ 50% in 76 (32%) cases. PD-L1-positive rate was 92% in squamous cell carcinomas and 67% in adenocarcinomas. PD-L1 Tumor Proportion Score was higher in squamous cell carcinomas (p = 0.004) and lower in adenocarcinomas (p = 0.003). Of the 196 non-squamous carcinomas, 35% had rat sarcoma viral oncogene homolog (RAS) mutations, 13% had epidermal growth factor receptor (EGFR) mutations, 2% had anaplastic lymphoma kinase (ALK) translocations and 2% had ROS1 translocations. Tumor Proportion Score ≥ 50% was seen in 34% (23/68), 28% (7/25) and 25% (1/4) of RAS, EGFR mutant, and ALK translocated carcinomas, respectively. There was no significant correlation between PD-L1 expression and molecular or genetic abnormalities, or other parameters including age, gender, stage, and smoking status. In our patient cohort, PD-L1 Tumor Proportion Score was significantly higher in squamous cell carcinomas and lower in adenocarcinomas. The overall prevalence of Tumor Proportion Score ≥ 50% is consistent with that reported in clinical trials.
Subject(s)
B7-H1 Antigen/biosynthesis , Biomarkers, Tumor/analysis , Carcinoma, Non-Small-Cell Lung/pathology , Lung Neoplasms/pathology , Adult , Aged , Aged, 80 and over , Australia , Female , Humans , Male , Middle AgedABSTRACT
AIMS: Micropapillary carcinomas, or carcinomas with a micropapillary component, are well recognised in the breast and other anatomical sites. However, they have seldom been described in the cervix. In this article, we present a clinicopathological analysis of eight cervical tumours that showed at least a focal (≥5%) component of micropapillary carcinoma. METHODS AND RESULTS: The study group comprised eight cervical carcinomas (four adenocarcinomas and four adenosquamous carcinomas) with a micropapillary component. The median patient age was 41.5 years (range 27-65 years). At presentation, five patients were stage IB, two were stage IIB, and one was stage IV. The micropapillary component accounted for ≤25% of the tumour on initial biopsy or resection specimens in all but one case. Immunohistochemistry showed 'inside-out' (reverse polarity) mucin 1 staining along the cell membrane abutting the stroma. Four patients developed metastasis, all of whom showed a pure micropapillary pattern; this led to a misdiagnosis of an apparently independent peritoneal serous carcinoma in one case. All tumours showed diffuse p16 expression, and all three cases that were tested were positive for human papillomavirus (HPV) 18. Three of the six patients with at least 12 months of follow-up died of disease, and one is alive with distant metastasis. CONCLUSIONS: Usual-type (HPV-related) cervical carcinomas may show micropapillary differentiation, usually as a focal finding, and the cells show reverse polarity like similar tumours arising in other sites. Micropapillary cervical carcinoma appears to be a clinically aggressive malignancy, although this needs to be confirmed in larger studies.
Subject(s)
Carcinoma, Papillary/pathology , Uterine Cervical Neoplasms/pathology , Adult , Aged , Carcinoma, Papillary/virology , Female , Humans , Middle Aged , Papillomavirus Infections/complications , Papillomavirus Infections/epidemiology , Uterine Cervical Neoplasms/virologyABSTRACT
The application of molecular genomics and our understanding of its clinical implications in the diagnosis, prognostication and treatment of lymphoproliferative disorders has rapidly evolved over the past few years. Of particular importance are indolent B-cell malignancies where tumour cell survival and proliferation are commonly driven by mutations involving the B-cell receptor and downstream signalling pathways. In addition, the increasing number of novel therapies and targeted agents have provided clinicians with new therapeutic options with the aim of exploiting such mutations. In this case report, we highlight one such success story involving the diagnostic impact of the MYD88L265P mutation in Waldenstrom's macroglobulinemia (WM), its prognostic implications and effect on choice of therapy in the era of novel therapies.
Subject(s)
Waldenstrom Macroglobulinemia/diagnosis , Agammaglobulinaemia Tyrosine Kinase , Aged, 80 and over , Humans , Immunoglobulin M/metabolism , Mutation/genetics , Myeloid Differentiation Factor 88/genetics , Protein-Tyrosine Kinases/genetics , Waldenstrom Macroglobulinemia/metabolism , Waldenstrom Macroglobulinemia/pathologyABSTRACT
BACKGROUND: The tall cell subtype of papillary thyroid cancer (TCPTC) is the most common aggressive subtype and often treated aggressively. This approach may not be necessary in smaller tumours without adverse histological characteristics. METHODS: 97 patients with TCPTC defined as a height-to-width ratio of ≥3:1 and at least 30% tall cells were compared against 390 classical papillary thyroid carcinoma (CPTC) based on tumour size with recurrence free survival (RFS) as the primary outcome. RESULTS: TCPTC are more likely to present with adverse histological characteristics. In smaller tumours (<2 âcm), only central lymph node metastasis (HR7.16 p â= â0.03) and multifocality (HR10.11 p â= â0.026) increased recurrence risk. In larger tumours, TCPTC histology (HR3.78 p â= â0.002), lymphovascular invasion (HR3.02 p â= â0.014) and central lymph node metastasis (HR3.24 p â< â0.001) significantly increased recurrence risk. CONCLUSION: TCPTC tumours <2 âcm without central lymph node metastasis and multifocality are similar in risk of recurrence to classical PTC and could be managed with lobectomy.
Subject(s)
Neoplasm Recurrence, Local , Thyroid Cancer, Papillary , Thyroid Neoplasms , Humans , Neoplasm Recurrence, Local/epidemiology , Neoplasm Recurrence, Local/pathology , Female , Male , Thyroid Neoplasms/pathology , Thyroid Neoplasms/surgery , Middle Aged , Thyroid Cancer, Papillary/pathology , Thyroid Cancer, Papillary/surgery , Thyroid Cancer, Papillary/mortality , Adult , Thyroidectomy , Lymphatic Metastasis , Retrospective Studies , Aged , Carcinoma, Papillary/pathology , Carcinoma, Papillary/surgery , Tumor Burden , Risk Assessment , Risk FactorsABSTRACT
Laparoscopy is currently considered to be the gold standard investigation in patients suspected to have endometriosis, but this is an invasive and relatively costly procedure and there may be significant delays in diagnosis. As the eutopic endometrium is recognized to be abnormal in patients with endometriosis, it has been suggested that endometrial sampling could provide an indirect diagnostic approach. In particular, recent reports have suggested that the presence of nerve fibers within the endometrial functional layer could represent a specific and sensitive marker of concurrent peritoneal endometriosis. However, such studies have been performed in select patient groups and using novel sampling and analytic techniques that are not used routinely in clinical pathology laboratories. The present study was performed upon conventional endometrial biopsies from 68 patients who underwent laparoscopy for suspected endometriosis. The biopsies were stained immunohistochemically for the neural marker PGP 9.5 and examined in a blinded manner. Endometrial functional layer nerve fibers were identified in 15 (22%) biopsies overall including 9/47 (19%) cases with histologically confirmed peritoneal endometriosis and 6/21 (29% cases) without endometriosis. There was no correlation between the presence of functional layer nerve fibers and the presenting symptoms, endometrial histology, or current hormonal therapy. In our experience, endometrial functional layer nerve fibers assessment performed using standard immunohistochemical techniques on routine biopsy specimens proved neither sensitive nor specific for the diagnosis of endometriosis. Pathologists and gynecologists considering this diagnostic approach should carefully consider the methodological factors that may influence its reliability.
Subject(s)
Endometriosis/diagnosis , Endometrium/innervation , Nerve Fibers/pathology , Adult , Biomarkers , Biopsy , Endometriosis/pathology , Endometrium/chemistry , Endometrium/pathology , Female , Humans , Immunohistochemistry , Laparoscopy , Middle Aged , Peritoneum/chemistry , Peritoneum/pathology , Sensitivity and Specificity , Ubiquitin Thiolesterase/analysisABSTRACT
Adenocarcinoma, not otherwise specified (NOS) is a heterogenous group of salivary gland tumors that likely contains distinct tumors that have not yet been characterized. Indeed, in recent years, cases previously diagnosed as adenocarcinoma, NOS have been recategorized into novel tumor designations such as secretory carcinoma, microsecretory adenocarcinoma, and sclerosing microcystic adenocarcinoma. We sought to describe a distinctive, hitherto-undescribed salivary gland tumor encountered in the authors' practices. Cases were pulled from the surgical pathology archives of the authors' institutions. Histologic, immunohistochemical, and clinical findings were tabulated, and targeted next-generation sequencing was performed on all cases. Nine cases were identified, arising in 8 women and 1 man ranging from 45 to 74 years (mean, 56.7 y). Seven tumors (78%) arose in the sublingual gland, while 2 (22%) arose in the submandibular gland. The cases shared a distinctive morphologic appearance. They were biphasic, with ducts scattered among a predominant polygonal cell with round nuclei, prominent nucleoli, and pale eosinophilic cytoplasm. These cells were arranged as trabeculae and palisaded as pseudorosettes around hyalinized stroma and vessels, resembling a neuroendocrine tumor. Four of the cases were well-circumscribed, while the remaining 5 showed infiltrative growth including perineural invasion in 2 (22%) and lymphovascular invasion in 1 (11%). Mitotic rates were low (mean, 2.2/10 HPFs); necrosis was absent. By immunohistochemistry, the predominant cell type was strongly positive for CD56 (9 of 9) and variably positive for pan-cytokeratin (AE1/AE3) (7 of 9) with patchy S100 (4 of 9), but negative for synaptophysin (0 of 9) and chromogranin (0 of 9), while the ducts were strongly positive for pan-cytokeratin (AE1/AE3) (9 of 9) and CK5/6 (7 of 7). Next-generation sequencing did not reveal any fusions or obvious driver mutations. All cases were resected surgically, with external beam radiation also done in 1 case. Follow-up was available in 8 cases; there were no metastases or recurrences after 4 to 160 months (mean, 53.1 mo). A dual population of scattered ducts with a predominance of CD56-positive neuroendocrine-like cells characterizes a unique salivary gland tumor which is often encountered in the sublingual glands of women, for which we propose the term "palisading adenocarcinoma." Although the tumor was biphasic and had a neuroendocrine-like appearance, it lacked convincing immunohistochemical evidence of myoepithelial or neuroendocrine differentiation. Although a subset showed unequivocally invasive growth, this tumor appears to behave in an indolent manner. Moving forward, recognition of palisading adenocarcinoma and its separation from other salivary adenocarcinomas, NOS will facilitate a better understanding of the characteristics of this previously unrecognized tumor.
Subject(s)
Adenocarcinoma , Carcinoma , Salivary Gland Neoplasms , Male , Humans , Female , Sublingual Gland/pathology , Salivary Gland Neoplasms/pathology , Adenocarcinoma/genetics , Adenocarcinoma/pathology , Immunohistochemistry , Biomarkers, Tumor/geneticsABSTRACT
Complete biomarker workup of non-small cell lung cancer (NSCLC) specimens is essential for appropriate and timely clinical management decisions. This can be challenging to achieve from small cytology and histology specimens, with increasing numbers of molecular and immunohistochemical biomarkers required. We conducted a 5 year retrospective audit of cases at our institution to assess the diagnostic and biomarker testing adequacy rates, particularly those specimens obtained with rapid onsite evaluation (ROSE), performed by a cytopathologist and a cytology scientist or pathology trainee, including all endobronchial ultrasound guided transbronchial needle aspirations (EBUS-TBNA), CT guided lung fine needle aspirations (FNA) and CT guided lung core biopsies. A total of 5,354 cases were identified, of which 92.2% had sufficient material for diagnosis. Of the 1506 cases identified with a recorded diagnosis of lung adenocarcinoma or NSCLC, not otherwise specified, 1001 (66.5%) had biomarker testing requested. Sufficient material was available in 89.5% of cases for a complete biomarker workup which included EGFR and KRAS mutational testing (all cases), ALK, ROS1 and PD-L1 immunohistochemistry (all cases), and ALK and ROS1 FISH (as required). For EGFR and KRAS mutational testing across both cytology and histology specimens, 99% of cases were sufficient. Of the samples in which a complete biomarker workup was unable to be performed, approximately half were only insufficient due to inadequate numbers of tumour cells for PD-L1 immunohistochemistry. Excluding PD-L1 IHC, 952 (95.1%) of samples obtained with ROSE were sufficient for the remainder of the testing requirements. Next generation sequencing using a 33 gene custom AmpliSeq panel was achieved in up to 72% of cases. In conclusion, small cytology and histology specimens obtained with ROSE are suitable for predictive biomarker testing in NSCLC, although attention needs to be paid to obtaining sufficient cells (>100) for PD-L1 immunohistochemistry.
Subject(s)
Carcinoma, Non-Small-Cell Lung , Lung Neoplasms , Humans , Carcinoma, Non-Small-Cell Lung/pathology , Lung Neoplasms/diagnosis , Lung Neoplasms/genetics , Lung Neoplasms/pathology , B7-H1 Antigen/metabolism , Retrospective Studies , Protein-Tyrosine Kinases , Proto-Oncogene Proteins p21(ras) , Proto-Oncogene Proteins , Endoscopic Ultrasound-Guided Fine Needle Aspiration , Biomarkers, Tumor , ErbB Receptors , Receptor Protein-Tyrosine KinasesABSTRACT
Recurrences frequently occur following surgical removal of primary tumors. In many cancers, adjuvant therapies have limited efficacy. Surgery provides access to the tumor microenvironment, creating an opportunity for local therapy, in particular immunotherapy, which can induce local and systemic anti-cancer effects. Here, we develop a surgically optimized biodegradable hyaluronic acid-based hydrogel for sustained intraoperative delivery of Toll-like receptor 3 agonist poly(I:C) and demonstrate that it significantly reduces tumor recurrence after surgery in multiple mouse models. Mechanistically, poly(I:C) induces a transient interferon alpha (IFNα) response, reshaping the tumor/wound microenvironment by attracting inflammatory monocytes and depleting regulatory T cells. We demonstrate that a pre-existing IFN signature predicts response to the poly(I:C) hydrogel, which sensitizes tumors to immune checkpoint therapy. The safety, immunogenicity, and surgical feasibility are confirmed in a veterinary trial in canine soft tissue tumors. The surgically optimized poly(I:C)-loaded hydrogel provides a safe and effective approach to prevent cancer recurrence.
Subject(s)
Hydrogels , Neoplasm Recurrence, Local , Mice , Animals , Dogs , Hydrogels/therapeutic use , Neoplasm Recurrence, Local/prevention & control , Immunotherapy , Disease Models, Animal , Tumor MicroenvironmentABSTRACT
An investigator-initiated, Australia-wide multi-centre retrospective observational study was undertaken to investigate the real-world prevalence of programmed death ligand-1 (PD-L1) expression in non-small cell lung carcinoma (NSCLC). Multiple centres around Australia performing PD-L1 immunohistochemistry (IHC) were invited to participate. Histologically confirmed NSCLC of any stage with a PD-L1 IHC test performed for persons aged ≥18 years between 1 January 2018 and 1 January 2020, and eligible for review, were identified at each centre, followed by data extraction and de-identification, after which data were submitted to a central site for collation and analysis. In total data from 6690 eligible PD-L1 IHC tests from histologically (75%) or cytologically (24%) confirmed NSCLC of any stage were reviewed from persons with a median age of 70 years, 43% of which were female. The majority (81%) of tests were performed using the PD-L1 IHC SP263 antibody with the Ventana BenchMark Ultra platform and 19% were performed using Dako PD-L1 IHC 22C3 pharmDx assay. Reported PD-L1 tumour proportion score (TPS) was ≥50% for 30% of all tests, with 62% and 38% scoring PD-L1 ≥1% and <1%, respectively. Relative prevalence of clinicopathological features with PD-L1 scores dichotomised to <50% and ≥50%, or to <1% and ≥1%, were examined. Females scored ≥1% slightly more often than males (64% vs 61%, respectively, p=0.013). However, there was no difference between sexes or age groups (<70 or ≥70 years) where PD-L1 scored ≥50%. Specimens from patients with higher stage (III/IV) scored ≥1% or ≥50% marginally more often compared to specimens from patients with lower stage (I/II) (p≤0.002). Proportions of primary and metastatic specimens did not differ where PD-L1 TPS was ≥1%, however more metastatic samples scored TPS ≥50% than primary samples (metastatic vs primary; 34% vs 27%, p<0.001). Cytology and biopsy specimens were equally reported, at 63% of specimens, to score TPS ≥1%, whereas cytology samples scored TPS ≥50% slightly more often than biopsy samples (34% vs 30%, respectively, p=0.004). Resection specimens (16% of samples tested) were reported to score TPS ≥50% or ≥1% less often than either biopsy or cytology samples (p<0.001). There was no difference in the proportion of tests with TPS ≥1% between PD-L1 IHC assays used, however the proportion of tests scored at TPS ≥50% was marginally higher for 22C3 compared to SP263 (34% vs 29%, respectively, p<0.001). These real-world Australian data are comparable to some previously published global real-world data, with some differences noted.
Subject(s)
B7-H1 Antigen , Carcinoma, Non-Small-Cell Lung , Lung Neoplasms , Adolescent , Adult , Aged , Female , Humans , Male , Australia/epidemiology , B7-H1 Antigen/metabolism , Biomarkers, Tumor/analysis , Carcinoma, Non-Small-Cell Lung/epidemiology , Carcinoma, Non-Small-Cell Lung/metabolism , Lung Neoplasms/epidemiology , Lung Neoplasms/metabolism , PrevalenceABSTRACT
PD-L1 immunohistochemistry has been approved as a diagnostic assay for immunotherapy. However, an international comparison across multiple cancers is lacking. This study aimed to assess the performance of PD-L1 diagnostic assays in non-small cell lung cancer (NSCLC), head and neck squamous cell cancer (HNSCC) and urothelial cancer (UC). The excisional specimens of NSCLC, HNSCC and UC were assayed by Ventana SP263 and scored at three sites in each country, including Australia, Brazil, Korea, Mexico, Russia and Taiwan. All slides were rotated to two other sites for interobserver scoring. The same cohort of NSCLC was assessed with Dako 22C3 pharmDx PD-L1 for comparison. The PD-L1 immunopositivity was scored according to the approved PD-L1 scoring algorithms which were the percentage of PD-L1-expressing tumour cell (TC) and tumour proportion score (TPS) by Ventana SP263 and Dako 22C3 staining, respectively. In NSCLC, the comparison demonstrated the comparability of the SP263 and 22C3 assays (cut-off of 1%, κ=0.71; 25%, κ=0.75; 50%, κ=0.81). The interobserver comparisons showed moderate to almost perfect agreement for SP263 in TC staining at 25% cut-off (NSCLC, κ=0.72 to 0.86; HNSCC, κ=0.60 to 0.82; UC, κ=0.68 to 0.91) and at 50% cut-off for NSCLC (κ=0.64 to 0.90). Regarding the immune cell (IC) scoring in UC, there was a lower correlation (concordance correlation coefficient=0.10 to 0.68) and poor to substantial agreements at the 1%, 5%, 10% and 25% cut-offs (κ= -0.04 to 0.76). The interchangeability of SP263 and 22C3 in NSCLC might be acceptable, especially at the 50% cut-off. In HNSCC, the performance of SP263 is comparable across five countries. In UC, there was low concordance of IC staining, which may affect treatment decisions. Overall, the study showed the reliability and reproducibility of SP263 in NSCLC, HNSCC and UC.
Subject(s)
Carcinoma, Non-Small-Cell Lung , Carcinoma, Transitional Cell , Head and Neck Neoplasms , Lung Neoplasms , Neoplasms, Squamous Cell , Urinary Bladder Neoplasms , Humans , Carcinoma, Non-Small-Cell Lung/diagnosis , Carcinoma, Non-Small-Cell Lung/pathology , Squamous Cell Carcinoma of Head and Neck/diagnosis , Reproducibility of Results , Lung Neoplasms/diagnosis , Lung Neoplasms/pathology , B7-H1 Antigen , Immunohistochemistry , Urinary Bladder Neoplasms/pathology , Head and Neck Neoplasms/diagnosis , Biomarkers, TumorABSTRACT
Mesothelioma is a cancer that typically originates in the pleura of the lungs. It rapidly invades the surrounding tissues, causing pain and shortness of breath. We compared cell lines injected either subcutaneously or intrapleurally and found that only the latter resulted in invasive and rapid growth. Pleural tumors displayed a transcriptional signature consistent with increased activity of nuclear receptors PPARα and PPARγ and with an increased abundance of endogenous PPAR-activating ligands. We found that chemical probe GW6471 is a potent, dual PPARα/γ antagonist with anti-invasive and anti-proliferative activity in vitro. However, administration of GW6471 at doses that provided sustained plasma exposure levels sufficient for inhibition of PPARα/γ transcriptional activity did not result in significant anti-mesothelioma activity in mice. Lastly, we demonstrate that the in vitro anti-tumor effect of GW6471 is off-target. We conclude that dual PPARα/γ antagonism alone is not a viable treatment modality for mesothelioma.
ABSTRACT
Low-coverage whole-genome sequencing (LC-WGS) can provide insight into oncogenic molecular changes. Serum extracellular vesicles (EV) represent a novel liquid biopsy source of tumoral DNA. This study compared copy number alteration (CNA) profiles generated from LC-WGS of formalin-fixed paraffin-embedded (FFPE) tumoral DNA and EV-DNA obtained from cancer patients. Patients with squamous cell carcinoma of the base of tongue (n = 3) and cutaneous squamous cell carcinoma (n = 2) were included. LC-WGS (0.5-1X coverage) was performed on FFPE-DNA and serum EV-DNA. Similarity between CNA profiles was analysed using QDNAseq. FFPE samples had a mean CNA of 31 (range 17-50) over 1.9 × 109 (range 1.0-2.6 × 109) bp in length, and EV samples had a mean CNA value of 17 (range 7-19) over 7.6 × 108 (range 2.9-15 × 108) bp in length. A mean of 8 (range 0-21) CNA over 5.9 × 108 (range 1.6-14 × 108) bp in length was found to overlap between EV and FFPE-derived samples per patient. Although the mean correlation efficient between samples was r = 0.34 (range - .08 to 0.99), this was not statistically significant (p > 0.05). Regions of highest deletion and duplication in FFPE samples were not well reflected in the EV-DNA. Selected CNA regions in EV-associated DNA were reflective of the primary tumor, however appreciation of global CNA and areas of most significant change was lost. The utility of LC-WGS of EV-derived DNA is likely limited to molecular alterations of known interest.
Subject(s)
Carcinoma, Squamous Cell/genetics , Extracellular Vesicles/genetics , Neoplasm Metastasis/genetics , Aged , Biomarkers, Tumor/blood , Biomarkers, Tumor/genetics , Carcinoma, Squamous Cell/metabolism , DNA Copy Number Variations/genetics , Extracellular Vesicles/pathology , Female , Formaldehyde/chemistry , Genome, Human/genetics , Genomics/methods , High-Throughput Nucleotide Sequencing/methods , Humans , Male , Middle Aged , Mouth Neoplasms/genetics , Mouth Neoplasms/metabolism , Neoplasm Metastasis/pathology , Neoplasms/genetics , Paraffin Embedding/methods , Tissue Fixation/methods , Whole Genome Sequencing/methodsABSTRACT
BACKGROUND AND AIM: Celiac disease (CD) and eosinophilic esophagitis (EE) are distinct disorders with specific clinico-pathological characteristics. Recent reports suggest an association between the 2. The aim of this study was to estimate the prevalence of EE among children diagnosed with CD in our institution in the last 8 years. MATERIALS AND METHODS: Princess Margaret Hospital in Western Australia is the state's only pediatric referral center and the Department of Anatomic Pathology handles almost all of the pediatric gastrointestinal biopsy specimens. All of the children who had histological confirmation of CD between January 2000 and November 2007 were identified. Among this cohort, those who had concurrent esophageal biopsies performed were obtained and those with histology consistent with EE identified. The slides of all of these cases were reviewed. Case notes of children with CD and EE were reviewed for demographic details, symptoms, endoscopic findings, and follow-up data. RESULTS: Among the total of 250 children diagnosed with CD during the study period, 121 had concurrent esophageal biopsies. Ten children had histological findings consistent with EE, although only 7 had endoscopic findings suggestive of EE. Median eosinophil count in these esophageal biopsies was 52 per high power field (range 23-80). Four children had follow-up endoscopies and all 4 demonstrated recovery of duodenal mucosa but persistent esophageal eosinophilia on gluten-free diet. In 3 children resolution of EE occurred after specific treatment of EE. CONCLUSIONS: The prevalence of EE in this cohort of children with CD is at least 4%. This is likely to be an underestimation because only 121 of 250 children had concurrent esophageal biopsies. Coexistent EE should be kept in mind in children undergoing endoscopy for suspected CD, and esophageal biopsies should be obtained, irrespective of whether esophageal mucosa appears normal or abnormal at endoscopy.
Subject(s)
Celiac Disease/complications , Eosinophilia/complications , Eosinophils/metabolism , Esophagitis/complications , Esophagus/pathology , Biopsy , Celiac Disease/pathology , Child , Child, Preschool , Diet, Gluten-Free , Duodenum/pathology , Eosinophilia/epidemiology , Eosinophilia/pathology , Esophagitis/epidemiology , Esophagitis/pathology , Female , Humans , Intestinal Mucosa/pathology , Male , PrevalenceABSTRACT
Increasing evidence points to the presence of low-level de novo T790M mutations in patients with non-small cell lung carcinoma (NSCLC) harboring activating EGFR mutations. We utilized digital PCR (dPCR), a highly sensitive gene mutation detection method, to detect pre-treatment T790M mutations in NSCLC tumor samples and correlated the T790M status with clinical features and patient outcomes. DNA extracted from pre-treatment NSCLC tumor tissue with known activating EGFR mutations, diagnosed between October 2010 and May 2017 at PathWest laboratory, was used to perform targeted dPCR for quantitative detection of T790M mutations. T790M was detected in 42 of 109 pre-treatment samples (38.5%). Median variant allele frequency was 0.14% (range 0.02-28.5%). Overall response rate to first generation EGFR tyrosine kinase inhibitors (TKI) was 67% regardless of T790M status. The median progression free survival was 10.7 (IQR 5.6-19.9) versus 6.7 (IQR 3.5-20.8) months in T790M negative and positive patients respectively. T790M positivity correlated with increased rate of early disease progression. It also correlated with increased mortality (HR 3.1 95%CI 1.2-8.1, p = 0.022) in patients who did not respond to TKI treatment. We detected a significant rate of low-level pre-treatment T790M mutations in NSCLC using highly sensitive dPCR. Low-level pre-treatment T790M did not impact treatment response rate or overall survival, but was associated with increased rate of early progression on TKI therapy.
Subject(s)
Biomarkers, Tumor/genetics , Carcinoma, Non-Small-Cell Lung/mortality , Drug Resistance, Neoplasm/genetics , Lung Neoplasms/mortality , Mutation , Protein Kinase Inhibitors/therapeutic use , Aged , Carcinoma, Non-Small-Cell Lung/drug therapy , Carcinoma, Non-Small-Cell Lung/genetics , Carcinoma, Non-Small-Cell Lung/pathology , ErbB Receptors/genetics , Female , Follow-Up Studies , Humans , Lung Neoplasms/drug therapy , Lung Neoplasms/genetics , Lung Neoplasms/pathology , Male , Middle Aged , Molecular Targeted Therapy , Prognosis , Retrospective Studies , Survival RateABSTRACT
We investigated whether a unique immune response was instigated with the development of oral tongue squamous cell carcinomas (OTSCC), with/without nodal involvement, with/without recurrent metastatic disease, or within tumor involved nodes. One hundred and ten formalin-fixed paraffin-embedded samples were collected from a retrospective cohort of 67 OTSCC patients and 10 non-cancerous tongue samples. Targets including CD4, CD8, FOXP3, PD-L1, and PD-1 were analyzed by immunohistochemistry. The Nanostring PanCancer Immune Profiling Panel was used for gene expression profiling. Data were externally validated in the The Cancer Genome Atlas (TCGA) head and neck (HNSCC), melanoma and lung squamous cell carcinoma (LSCC) cohorts. A 24-immune gene signature was identified that discriminated more aggressive OTSCC cases, and although not prognostic in HNSCC was associated with survival in other TCGA cohorts (improved survival for melanoma, P < .001 and worse survival for LSCC, P = .038). OTSCC exhibited concordant gene and immunohistochemical (IHC) features characterized by a TH-2 biased, proinflammatory profile with upregulated B cell and neutrophil gene activity and increased CD4, FOXP3, and PD-L1 expression (P < .001 for all by IHC). Compared to less advanced disease, nodal involvement and recurrent OTSCC did not induce a different immune response although recurrent disease was characterized by significantly higher PD-L1 expression (P = .004 by SP263, P = .013 by 22C3, P = .004 for gene expression). Identification of a gene signature associated with different prognostic effects in other cancers highlights common pathways of immune dysregulation that are impacted by the tumor origin. The significant immunosuppressive signaling in OTSCC indicates primary failure of immune system to control carcinogenesis emphasizing the need for early, combination therapeutic approaches.
Subject(s)
Squamous Cell Carcinoma of Head and Neck/immunology , Tongue Neoplasms/immunology , Aged , B7-H1 Antigen/analysis , CD4 Antigens/analysis , CD8 Antigens/analysis , Female , Forkhead Transcription Factors/analysis , Gene Expression , Gene Expression Profiling , Humans , Lung Neoplasms/immunology , Lung Neoplasms/mortality , Lymph Nodes/pathology , Male , Melanoma/immunology , Melanoma/mortality , Middle Aged , Prognosis , Programmed Cell Death 1 Receptor/analysis , Retrospective Studies , Squamous Cell Carcinoma of Head and Neck/genetics , Squamous Cell Carcinoma of Head and Neck/mortality , Squamous Cell Carcinoma of Head and Neck/pathology , Tongue/immunology , Tongue Neoplasms/genetics , Tongue Neoplasms/mortality , Tongue Neoplasms/pathologyABSTRACT
PURPOSE: This study compares the detection sensitivity of two separate liquid biopsy sources, cell-free (cf) DNA/RNA and extracellular vesicle (EV)-associated DNA/RNA (EV-DNA/RNA), to identify circulating Human Papilloma Virus (HPV) DNA/RNA in plasma obtained from patients with oropharyngeal squamous cell carcinoma (OPCSCC). We also report on the longitudinal changes observed in HPV-DNA levels in response to treatment. EXPERIMENTAL DESIGN: A prospective study was conducted that included 22 patients with locally advanced disease and six patients with metastatic OPCSCC. Twenty-three patients had HPV-related OPCSCC defined by p16 immunohistochemistry. Levels of circulating HPV-DNA and HPV-RNA from plasma-derived cf-DNA/RNA and EV-DNA/RNA were quantified using digital droplet PCR. RESULTS: Circulating HPV-DNA was detected with higher sensitivity in cf-DNA compared to EV-DNA at 91% vs. 42% (p = <0.001). Similarly, circulating tumoral HPV-RNA was detected at a higher sensitivity in cf-RNA compared to EV-RNA, at 83% vs. 50% (p = 0.0019). In the locally advanced cohort, 100% (n = 16) of HPV-OPCSCC patients demonstrated a reduction in circulating HPV-DNA levels in cf-DNA following curative treatment, with 81% of patients demonstrating complete clearance to undetectable levels. However, in metastatic HPV-OPCSCC patients (n = 4), HPV-DNA levels did not correlate with treatment response. CONCLUSION: Our study demonstrates that although HPV-DNA/RNA can be detected in EV associated DNA/RNA, cf-DNA/RNA is the more sensitive liquid biopsy medium. As circulating HPV-DNA levels were found to only correlate with treatment response in the locally advanced but not metastatic setting in our small cohort of patients, the use of HPV-DNA as a dynamic biomarker to monitor treatment response requires further evaluation.
Subject(s)
Carcinoma, Squamous Cell/pathology , Cell-Free Nucleic Acids/analysis , DNA, Viral/analysis , Extracellular Vesicles/virology , Human Papillomavirus DNA Tests/methods , Oropharyngeal Neoplasms/pathology , Aged , Carcinoma, Squamous Cell/blood , Carcinoma, Squamous Cell/virology , Cell-Free Nucleic Acids/genetics , DNA, Viral/genetics , Female , Human Papillomavirus DNA Tests/standards , Humans , Limit of Detection , Liquid Biopsy/methods , Liquid Biopsy/standards , Male , Middle Aged , Oropharyngeal Neoplasms/blood , Oropharyngeal Neoplasms/virologyABSTRACT
There is increasing interest in p53 immunohistochemistry as an adjunct to haematoxylin and eosin (H&E) assessment for dysplasia in oesophageal Barrett's mucosa; however, published information on the patterns of staining remains scant. Here, we present descriptions of normal and aberrant p53 staining in non-neoplastic and dysplastic Barrett's mucosa in endoscopic mucosal resections. A retrospective series of archival endoscopic mucosal resections for biopsy proven dysplasia at our institution were retrieved for this study, comprising 28 sections from 23 patients. p53 immunohistochemistry was performed using an in-house optimised protocol and the staining pattern assessed in H&E confirmed non-neoplastic, dysplastic and neoplastic areas of Barrett's mucosa with regard to individual cell intensity and location of positive cells with respect to gland microanatomy. In non-neoplastic epithelium, normal p53 staining was weak, heterogenous and localised to the crypts. In dysplastic epithelium, p53 over-expression was seen which was of moderate to strong intensity in either a crypt predominant location or diffuse involving crypt and surface epithelium. The crypt predominant pattern was observed more commonly in low grade dysplasia while the diffuse pattern was more commonly seen in high grade dysplasia. In a minority of cases, there was complete loss of p53 staining in dysplastic epithelium and contiguous neoplasia (null phenotype). p53 immuno-expression in non-neoplastic and dysplastic Barrett's mucosa is distinctive when interpreted with regard to cell intensity and gland microanatomy. We propose that these staining patterns may assist in the interpretation of dysplasia in endoscopic biopsies of Barrett's mucosa.
Subject(s)
Barrett Esophagus/metabolism , Esophageal Mucosa/metabolism , Esophageal Neoplasms/metabolism , Tumor Suppressor Protein p53/metabolism , Barrett Esophagus/pathology , Endoscopic Mucosal Resection , Esophageal Mucosa/pathology , Esophageal Neoplasms/pathology , Humans , Immunohistochemistry , Retrospective StudiesABSTRACT
OBJECTIVES: To describe three further cases of anaplastic large cell lymphoma (ALCL) occurring in patients with preexisting chronic lymphocytic leukemia (CLL). We also reviewed the literature of previously published cases. METHODS: We discuss the clinical features, histopathology, and outcomes for three patients with ALCL and CLL from Perth, Australia. The cases were also included in a literature review of existing cases and comparisons were made with our cohort. RESULTS: The three patients included two men (aged 77 and 74 years) and one woman (aged 66 years). All had a history of untreated CLL with diagnosis established 4 to 16 years before. They had lymphadenopathy and/or cutaneous/soft tissue lesions that proved to be ALCL, ALK+ (one case) or ALCL, ALK- (two cases). CONCLUSIONS: Further research is required in this area to establish prognostic and management recommendations. Increasing numbers of cases are being described. Positron emission tomography with computed tomography was not useful in our cohort for diagnosing progression.
Subject(s)
Leukemia, Lymphocytic, Chronic, B-Cell/pathology , Lymphoma, Large-Cell, Anaplastic/pathology , Neoplasms, Multiple Primary/pathology , Aged , Female , Humans , MaleABSTRACT
Cancer immunotherapy using antibodies that target immune checkpoints has delivered outstanding results. However, responses only occur in a subset of patients, and it is not fully understood what biological processes determine an effective outcome. This lack of understanding hinders the development of rational combination treatments. We set out to define the pretreatment microenvironment associated with an effective outcome by using the fact that inbred mouse strains bearing monoclonal cancer cell line-derived tumors respond in a dichotomous manner to immune checkpoint blockade (ICB). We compared the cellular composition and gene expression profiles of responsive and nonresponsive tumors from mice before ICB and validated the findings in cohorts of patients with cancer treated with ICB antibodies. We found that responsive tumors were characterized by an inflammatory gene expression signature consistent with up-regulation of signal transducer and activator of transcription 1 (STAT1) and Toll-like receptor 3 (TLR3) signaling and down-regulation of interleukin-10 (IL-10) signaling. In addition, responsive tumors had more infiltrating-activated natural killer (NK) cells, which were necessary for response. Pretreatment of mice with large established tumors using the STAT1-activating cytokine interferon-γ (IFNγ), the TLR3 ligand poly(I:C), and an anti-IL-10 antibody sensitized tumors to ICB by attracting IFNγ-producing NK cells into the tumor, resulting in increased cure rates. Our results identify a pretreatment tumor microenvironment that predicts response to ICB, which can be therapeutically attained. These data suggest a biomarker-driven approach to patient management to establish whether a patient would benefit from treatment with sensitizing therapeutics before ICB.