ABSTRACT
Medullary carcinoma of the colon is a unique histologic subtype of microsatellite unstable colorectal carcinoma but little is known regarding its tumor-immunoregulatory microenvironment. The aims of this study were to characterize the immune environment of medullary carcinoma and compare it with other microsatellite unstable and microsatellite stable colorectal carcinomas. An initial gene expression microarray analysis of six cases of medullary carcinoma was used to detect potentially differentially expressed genes. We extended this analysis utilizing genomic data from the Cancer Genome Atlas to compare eight cases of medullary carcinoma with other microsatellite unstable and stable carcinomas. Finally, we evaluated expression of key immune pathway proteins and lymphocyte subsets via immunohistochemistry of a large group of medullary carcinomas (n=105) and compared these findings with three other groups: poorly differentiated, microsatellite unstable well-differentiated and microsatellite stable well-differentiated carcinomas. Microarray and the Cancer Genome Atlas data analysis identified significant upregulation of several immunoregulatory genes induced by IFNγ including IDO-1, WARS (tRNA(trp)), GBP1, GBP4, GBP5, PDCD1 (PD-1), and CD274 (PD-L1) in medullary carcinoma compared with other microsatellite unstable and microsatellite stable tumors. By immunohistochemistry, IDO-1 was expressed in 64% of medullary carcinomas compared with 19% (9/47) of poorly differentiated carcinomas, 14% (3/22) of microsatellite unstable, and 7% (2/30) of the microsatellite stable well-differentiated carcinomas (P<0.0001). tRNA(trp) was overexpressed in 81% (84/104) of medullary carcinomas, 19% (9/47) of poorly differentiated, 32% (7/22) of microsatellite unstable, and 3% (1/30) of microsatellite stable well-differentiated carcinomas (P<0.0001). Medullary carcinoma had higher mean CD8+ and PD-L1+ tumor-infiltrating lymphocytes compared with all other groups (P<0.0001). This study demonstrates overexpression of several immunoregulatory genes in microsatellite unstable colorectal carcinomas and that expression of these genes and proteins is more prevalent in the medullary carcinoma subtype, which may be of use both diagnostically and therapeutically.
Subject(s)
Carcinoma, Medullary/genetics , Carcinoma, Medullary/pathology , Colonic Neoplasms/genetics , Colonic Neoplasms/pathology , Tumor Microenvironment/immunology , Adult , Aged , Aged, 80 and over , Carcinoma, Medullary/immunology , Colonic Neoplasms/immunology , Female , Gene Expression Profiling , Humans , Lymphocytes, Tumor-Infiltrating/pathology , Male , Middle Aged , TranscriptomeABSTRACT
INTRODUCTION: Different pathological types of colorectal cancer have distinguished immune landscape, and the efficacy of immunotherapy will be completely different. Colorectal medullary carcinoma, accounting for 2.2-3.2%, is characterized by massive lymphocyte infiltration. However, the attention to the immune characteristics of colorectal medullary carcinoma is insufficient. AREA COVERED: We searched the literature about colorectal medullary carcinoma on PubMed through November 2023to investigate the hallmarks of colorectal medullary carcinoma's immune landscape, compare medullary carcinoma originating from different organs and provide theoretical evidence for precise treatment, including applying immunotherapy and BRAF inhibitors. EXPERT OPINION: Colorectal medullary carcinoma is a pathological subtype with intense immune response, with six immune characteristics and has the potential to benefit from immunotherapy. Mismatch repair deficiency, ARID1A missing and BRAF V600E mutation often occurs. IFN-γ pathway is activated and PD-L1 expression is increased. Abundant lymphocyte infiltration performs tumor killing function. In addition, BRAF mutation plays an important role in the occurrence and development, and we can consider the combination of BRAF inhibitors and immunotherapy in patients with BRAF mutant. The exploration of colorectal medullary carcinoma will arouse researchers' attention to the correlation between pathological subtypes and immune response, and promote the process of precise immunotherapy.
Subject(s)
Carcinoma, Medullary , Colorectal Neoplasms , Immune Checkpoint Inhibitors , Mutation , Proto-Oncogene Proteins B-raf , Humans , Immune Checkpoint Inhibitors/therapeutic use , Colorectal Neoplasms/immunology , Colorectal Neoplasms/pathology , Colorectal Neoplasms/drug therapy , Colorectal Neoplasms/genetics , Colorectal Neoplasms/therapy , Proto-Oncogene Proteins B-raf/genetics , Proto-Oncogene Proteins B-raf/antagonists & inhibitors , Carcinoma, Medullary/immunology , Carcinoma, Medullary/genetics , Carcinoma, Medullary/pathology , Carcinoma, Medullary/therapy , Immunotherapy/methods , Lymphocytes, Tumor-Infiltrating/immunology , Animals , B7-H1 Antigen/antagonists & inhibitors , B7-H1 Antigen/metabolism , B7-H1 Antigen/immunologyABSTRACT
AIMS: Medullary breast cancer (MBC) is a biologically distinct subtype of breast cancer characterized by prominent lymphocytic infiltrates and a favourable clinical outcome. Tumour-infiltrating CD8+ effector T cells may contribute to the good prognosis of this type of cancer; however, certain subtypes of lymphocyte, such as FoxP3+ regulatory T cells (Tregs), can also suppress antitumour immunity. METHODS AND RESULTS: We determined tumour infiltration by FoxP3+, CCL22+ and CD8+ cells in paraffin-embedded sections of MBC, and, as a reference, in samples of grade 3 ductal, lobular and mucinous breast cancer. All analysed MBCs were strongly infiltrated by FoxP3+ cells, whereas only weak infiltrates were detected in ductal or lobular breast cancer. This finding was unexpected, given the good prognosis of MBC. Strikingly, the number of CD8+ T cells exceeded the number of FoxP3+ cells in MBC (ratio of CD8+ to FoxP3+ cells of 2.6), whereas equal amounts of both cell types were found in ductal breast cancer (ratio of CD8+ to FoxP3+ cells of 1.1). In both types of breast cancer, we also detected cells expressing the Treg-attracting chemokine CCL22. CONCLUSIONS: In breast cancer, a predominance of tumour-infiltrating CD8+ over FoxP3+ cells was observed in MBC. Thus, the ratio of CD8+ to FoxP3+ cells rather than the absolute number of intratumoral FoxP3+ cells may be predictive for the clinical outcome of cancer.
Subject(s)
Breast Neoplasms/pathology , CD8-Positive T-Lymphocytes/immunology , Carcinoma, Medullary/pathology , Forkhead Transcription Factors/immunology , Lymphocytes, Tumor-Infiltrating/immunology , T-Lymphocyte Subsets/immunology , Adult , Aged , Breast Neoplasms/immunology , Carcinoma, Medullary/immunology , Enzyme-Linked Immunosorbent Assay , Female , Humans , Immunohistochemistry , Middle Aged , Neoplasm StagingABSTRACT
AIM: To immunohistochemically evaluate the expression of MAGE-A1, MAGE-A, and NY-ESO-1 cancer/testis (C/T) tumor antigens in medullary breast cancer (MBC) tumor samples and to analyze it in relation to the clinicopathological features. METHODS: This retrospective study included samples from 49 patients: 40 with typical MBC and 9 with atypical MBC. Tumor specimens were obtained from patients operated on in the University Hospital for Tumors and the Sisters of Mercy University Hospital, Zagreb, Croatia, from 1999 to 2005. Standard immunohistochemistry was used on archival paraffin-embedded MBC tissues. RESULTS: MAGE-A1, MAGE-A, and NY-ESO-1 antigens were expressed in 33% (16/49), 33% (16/49), and 22% (11/49) of patients, respectively. No difference between the groups with and without C/T tumor antigen expression in age at diagnosis, tumor size, axillary lymph node metastasis, adjuvant therapy, and HER-2 expression was identified. Significantly more patients died in the MAGE-A-positive group than in the MAGE-A-negative group (P=0.010), whereas a borderline significance was found between MAGE-A1-positive and the MAGE-A1-negative group (P=0.079) and between NY-ESO-1-positive and NY-ESO-1-negative group (P=0.117). Overall survival, as evaluated by the Kaplan-Meier curves, was lower in MAGE-A1- (P=0.031), MAGE-A- (P=0.004), NY-ESO-1-positive groups (P=0.077). CONCLUSION: Expression of C/T antigens may represent a marker of potential prognostic relevance in MBC.
Subject(s)
Antigens, Neoplasm/analysis , Antigens/genetics , Antigens/immunology , Breast Neoplasms/immunology , Carcinoma, Medullary/immunology , Gene Expression Regulation, Neoplastic/immunology , Membrane Proteins/analysis , Neoplasm Proteins/analysis , Peptide Fragments/analysis , Adult , Aged , Aged, 80 and over , Antigens, Neoplasm/genetics , Antigens, Neoplasm/immunology , Croatia , Female , Humans , Immunohistochemistry , Membrane Proteins/genetics , Membrane Proteins/immunology , Middle Aged , Neoplasm Proteins/genetics , Neoplasm Proteins/immunology , Peptide Fragments/genetics , Peptide Fragments/immunology , Retrospective Studies , Survival AnalysisABSTRACT
OBJECTIVE: To investigate the expression of indoleamine 2, 3-dioxygenase (IDO) in breast cancer and its correlation with clinicopathologic factors and prognosis. METHODS: The expression of IDO, CD31, CD105 proteins in 40 specimens of breast cancer were assessed by immunohistochemistry. RESULTS: The overexpression rate of IDO in breast cancer was 67.5% (27/40), and expression of IDO was closely associated with clinical stage and lymph nodes metastasis. The disease-free survival rate in patients with IDO overexpression was not significantly lower than that in patients with negative or low expression of IDO (P > 0.05). Moreover, the expression of IDO was positively correlated with CD105-labeled microvessel density (r = 0.659, P < 0.05). CONCLUSIONS: Expression of IDO is associated with clinical stage and lymph nodes metastasis, and microvessel densitty. IDO expression may promote the growth and metastasis of breast cancer, probably via the increased agiogenesis. A larger sample study is needed to verify whether the prognosis of beast cancer is significantly correlated with IDO expression.
Subject(s)
Breast Neoplasms/enzymology , Carcinoma, Ductal, Breast/enzymology , Indoleamine-Pyrrole 2,3,-Dioxygenase/metabolism , Microvessels , Adenocarcinoma/enzymology , Adenocarcinoma/immunology , Adenocarcinoma/pathology , Adult , Aged , Antigens, CD/metabolism , Breast Neoplasms/immunology , Breast Neoplasms/pathology , Carcinoma, Ductal, Breast/immunology , Carcinoma, Ductal, Breast/pathology , Carcinoma, Medullary/enzymology , Carcinoma, Medullary/immunology , Carcinoma, Medullary/pathology , Disease-Free Survival , Endoglin , Female , Follow-Up Studies , Humans , Immunohistochemistry , Lymphatic Metastasis , Microvessels/enzymology , Microvessels/immunology , Middle Aged , Neoplasm Staging , Platelet Endothelial Cell Adhesion Molecule-1/metabolism , Receptors, Cell Surface/metabolism , Survival RateABSTRACT
Medullary carcinoma (MC) of the breast is a high grade carcinoma that has a relatively favourable prognosis compared to atypical medullary carcinoma (AMC) and other more common breast carcinomas. In a retrospective study in Brunei Darussalam of all available biopsy samples, we compared the nature of the tumour-infiltrating lymphocytes (TILs) in MC and AMC in relation to recorded tumour characteristics. CD4, CD8, CD20, CD25, CD45RO, and CD56 and common tumour biomarkers were detected immunohistochemically. The 11 cases of MC had no nodal metastases and survived without relapse, suggesting good tumour control. In contrast, 7 cases of nodal metastases and 1 relapse were observed in 12 AMCs. Although not statistically significant, there was a tendency for a greater proportion of AMCs to express the Her2/neu oncogene. Higher proportions of CD45RO+ and CD8+ cells, and lower levels of CD20+ cells, were characteristic of TILs in MC compared to AMC. The ratio of CTL to B-lineage cells in TILs in both tumours considered together was inversely related to the expression of HER2/neu and the presence of nodal metastases. The findings suggest that CTLs, rather than antibodies, may give better tumour control in MC relative to AMC. We propose that a comparison of the cellular, molecular and immunological characteristics of MC and AMC, as a paired model system, in a multi-centre investigation with a much larger number of samples will be valuable for better understanding mechanisms of tumour immunity.
Subject(s)
B-Lymphocytes/immunology , Breast Neoplasms/immunology , Carcinoma, Medullary/immunology , Lymphocytes, Tumor-Infiltrating/immunology , T-Lymphocytes, Cytotoxic/immunology , Adult , Aged , Antigens, CD/metabolism , Biomarkers, Tumor/analysis , Breast Neoplasms/genetics , Breast Neoplasms/pathology , Carcinoma, Medullary/genetics , Carcinoma, Medullary/pathology , Cell Lineage/immunology , Female , Genes, erbB-2 , Humans , Immunohistochemistry , Immunophenotyping , Middle Aged , Retrospective StudiesABSTRACT
BACKGROUND: Immunological response of the human body is controlled by the suppressive characteristics of regulatory T cells (Tregs). In various diseases a change in the number of Tregs is evident. For example, whereas Tregs are reduced in auto-immunological processes, an increase of Tregs is found with various malignant tumors. Regarding medullary thyroid carcinoma (MTC) no such studies have been performed to date. METHODS: Expression of CD4 and CD25 in CD45+ leukocytes from blood and lymph nodes was studied by flow cytometry in patients with MTC and patients with benign goiter. We also examined the marker forkhead box P3 (FoxP3), an intracellular transcription factor, which is supposed to be the most specific marker for Tregs. Immunohistochemical staining for FoxP3 was performed on lymph node and thyroid tissue. RESULTS: The number of FoxP3+ lymphocytes in peripheral blood was significantly higher in patients with MTC than in controls (p = 0.02). This result was confirmed immunohistochemically in lymph node and thyroid tissue, as well as in carcinoma tissue. No difference in CD4+CD25+ lymphocytes was observed between the two groups. After clinical staging (International Union against Cancer-UICC-stages) of MTC patients, triplication of FoxP3+ lymphocytes could be observed from MTC < UICC II to MTC > UICC II. CONCLUSIONS: An increase of FoxP3+ lymphocytes could be shown in peripheral blood of patients with MTC but not in patients with benign goiter; this increase also correlates with findings in lymph nodes and thyroid gland. The number of FoxP3+ cells correlated with the patients' prognosis. Therefore, FoxP3+ lymphocytes are a good diagnostic criterion for malignancy in patients with medullary thyroid carcinoma, and their presence at staging may influence therapeutic decisions.
Subject(s)
Carcinoma, Medullary/immunology , Lymph Nodes/immunology , T-Lymphocytes, Regulatory/immunology , Thyroid Neoplasms/immunology , Adult , Aged , CD4 Antigens/metabolism , Carcinoma, Medullary/metabolism , Carcinoma, Medullary/surgery , Female , Flow Cytometry , Forkhead Transcription Factors/metabolism , Humans , Immunohistochemistry , Interleukin-2 Receptor alpha Subunit/metabolism , Male , Middle Aged , T-Lymphocytes, Regulatory/metabolism , Thyroid Neoplasms/metabolism , Thyroid Neoplasms/surgeryABSTRACT
Renal medullary carcinoma (RMC) is a highly lethal malignancy that mainly afflicts young individuals of African descent and is resistant to all targeted agents used to treat other renal cell carcinomas. Comprehensive genomic and transcriptomic profiling of untreated primary RMC tissues was performed to elucidate the molecular landscape of these tumors. We found that RMC was characterized by high replication stress and an abundance of focal copy-number alterations associated with activation of the stimulator of the cyclic GMP-AMP synthase interferon genes (cGAS-STING) innate immune pathway. Replication stress conferred a therapeutic vulnerability to drugs targeting DNA-damage repair pathways. Elucidation of these previously unknown RMC hallmarks paves the way to new clinical trials for this rare but highly lethal malignancy.
Subject(s)
Biomarkers, Tumor/metabolism , Carcinoma, Medullary/pathology , Carcinoma, Renal Cell/pathology , Chromosome Aberrations , DNA Replication , Kidney Neoplasms/pathology , SMARCB1 Protein/metabolism , Adult , Animals , Apoptosis , Biomarkers, Tumor/genetics , Carcinoma, Medullary/genetics , Carcinoma, Medullary/immunology , Carcinoma, Renal Cell/genetics , Carcinoma, Renal Cell/immunology , Cell Proliferation , Cohort Studies , DNA Copy Number Variations , Female , Gene Expression Regulation, Neoplastic , Genomics , High-Throughput Nucleotide Sequencing , Humans , Kidney Neoplasms/genetics , Kidney Neoplasms/immunology , Male , Membrane Proteins/genetics , Membrane Proteins/metabolism , Mice , Mice, Nude , Nucleotidyltransferases/genetics , Nucleotidyltransferases/metabolism , Prognosis , Proto-Oncogene Proteins c-myc/genetics , Proto-Oncogene Proteins c-myc/metabolism , SMARCB1 Protein/genetics , Tumor Cells, Cultured , Xenograft Model Antitumor AssaysABSTRACT
Activating mutations in the Ret proto-oncogene are responsible for occurrence of multiple endocrine neoplasia (MEN) type 2A and 2B, and familial medullary thyroid carcinoma (FMTC). A striking genotype-phenotype correlation between the mutated RET codon and clinical manifestation implies that tumorigenesis is conditioned by the type of mutation. We investigated gene expression profiles between and within distinct MEN2 subtypes through whole-genome microarray analysis in tumors induced by NIH-3T3 cells transformed with defined RET-MEN2A (C609Y, C634R), MEN2B, (A883F, M918T), and FMTC (Y791F) mutations. Expression profiling identified a statistically significant modification of 1494 genes, 628 down- and 866 upregulated in MEN2B compared with MEN2A/FMTC tumors. By contrast, no obvious alterations were observed among individual MEN2B and MEN2A type mutations, or between MEN2A and FMTC. Functional clustering of differential genes revealed RET-MEN2B specific upregulation of genes associated with novel growth and survival pathways. Intriguingly, RET-MEN2A/FMTC-specific tumors were characterized by a considerable number of genes involved in the host antitumor immune response via stimulation of natural killer/T-cell proliferation, migration, and cytotoxicity, which were completely absent in RET-MEN2B related cancers. QPCR on tumors versus cultured NIH-RET cell lines demonstrated that they are largely attributed to the host innate immune system, whereas expression of CX3CL1 involved in leukocyte recruitment is exclusively RET-MEN2A/FMTC tumor cell dependent. In correlation, massive inflammatory infiltrates were apparent only in tumors carrying MEN type 2A/FMTC mutations, suggesting that RET-MEN2B receptors specifically counteract immune infiltration by preventing chemokine expression, which may contribute to the different clinical outcome of both subtypes.
Subject(s)
Carcinoma, Medullary/genetics , Gene Expression Profiling , Gene Expression Regulation, Neoplastic , Multiple Endocrine Neoplasia Type 2a/genetics , Multiple Endocrine Neoplasia Type 2b/genetics , Thyroid Neoplasms/genetics , Animals , Carcinoma, Medullary/immunology , Killer Cells, Natural/immunology , Mice , Multiple Endocrine Neoplasia Type 2a/immunology , Multiple Endocrine Neoplasia Type 2b/immunology , NIH 3T3 Cells , Oligonucleotide Array Sequence Analysis , Point Mutation , Proto-Oncogene Proteins c-ret/genetics , Reverse Transcriptase Polymerase Chain Reaction , Thyroid Neoplasms/immunologyABSTRACT
Immunotherapy with autologous dendritic cells (DCs) loaded with tumor lysate(s) from allogeneic tumor cell lines is a novel strategy to induce immune responses in cancer patients. We report on a pilot trial of autologous DCs pulsed with tumor cell lysate derived from allogeneic medullary thyroid carcinoma (MTC) cell lines in patients with metastatic MTC. The purpose of this study was to assess the safety, resulting immune responses and clinical activity of the DCs. DCs were injected into a groin lymph node at 3-week intervals. Monitoring included serial calcitonin tumor marker measurements, radiological imaging and immunological in vitro tests (T-cell interferon-gamma detection assay, T-cell cytotoxicity assay). Ten patients (median age 47 years, range 29-77) were enrolled. DC vaccinations were well-tolerated and safe. After a median follow-up of 11 months, (range 7-26), 3 (30%) of 10 patients had stable disease, while 7 (70%) of the patients progressed during treatment. In 2 patients with stable disease, calcitonin decreased below treatment levels, paralleled by a T-cell-mediated immune response. Notably, treatment with DCs pulsed with a combination of different tumor cell lysates was followed by a calcitonin decrease in 4 patients who had previously experienced a calcitonin increase during monotherapy with DCs pulsed with a single lysate. Allogeneic tumor cell lysate-based DC immunotherapy is well-tolerated and safe. Combined treatment with different tumor cell lysate-pulsed DCs increases the likelihood of a calcitonin tumor marker response and should therefore be preferred over monotherapy with DCs pulsed with a single lysate.
Subject(s)
Carcinoma, Medullary/therapy , Dendritic Cells/transplantation , Immunotherapy, Adoptive , Thyroid Neoplasms/therapy , Adult , Aged , Biomarkers, Tumor/blood , Calcitonin/blood , Cancer Vaccines , Carcinoma, Medullary/diagnosis , Carcinoma, Medullary/immunology , Carcinoma, Medullary/secondary , Cell Line, Tumor , Dendritic Cells/immunology , Disease Progression , Female , Humans , Immunotherapy, Adoptive/adverse effects , Interferon-gamma/metabolism , Male , Middle Aged , Pilot Projects , T-Lymphocytes, Cytotoxic/immunology , Thyroid Neoplasms/diagnosis , Thyroid Neoplasms/immunology , Thyroid Neoplasms/secondary , Time Factors , Transplantation, Autologous , Treatment OutcomeABSTRACT
Up to now, no relevant tumor antigen has been identified in medullary thyroid carcinoma (MTC). The aim of the present study was to prove the concept of an immunization with an amino acid-modified calcitonin (CT) for the treatment of MTC in a transgenic mouse model. Amino acid-modified (human) CT has been chosen for vaccination because of its higher binding affinity to the murine H2-Kb-MHC molecule. Mice were immunized over 6 months with monthly injections of amino acid-modified CT-pulsed dendritic cells. For enumeration of tumor epitope-specific CD8+ cytotoxic T cells, tetramer analyses were performed. CT peptide-treated mice revealed a mean 0.73 +/- 0.45 and 0.91 +/- 0.59% positive cells, depending on the two tetramers tested, whereas no increase was seen in control protein-immunized mice (0.08-0.12% tetramer-positive cells). Importantly, the subset of CT-specific CD8+ T cells also showed a high expression of interferon-gamma. In line with these results, CT-immunized mice also showed an intratumor infiltration with CD8+ T lymphocytes. Importantly, we also found a diminished tumor outgrowth of -57% and a decrease of the serum CT levels (2.0 +/- 0.1 pg/ml) compared with control protein-immunized Ret/Cal mice (3.0 +/- 0.4 pg/ml). In summary, we show that amino acid-modified CT is recognized from the immune system leading to a specific antitumor immune response and a diminished tumor outgrowth in transgenic MTC mice. The results are of potential importance because they might be applicable to patients with metastatic spread of a MTC.
Subject(s)
Antigens, Neoplasm/immunology , Calcitonin/immunology , Carcinoma, Medullary/therapy , Thyroid Neoplasms/therapy , Amino Acids/chemistry , Animals , Antibody Specificity , CD8-Positive T-Lymphocytes/immunology , Calcitonin/chemistry , Carcinoma, Medullary/immunology , Carcinoma, Medullary/pathology , Disease Models, Animal , Epitopes, T-Lymphocyte/immunology , Immunization/methods , Immunotherapy, Active , Mice , Mice, Inbred C57BL , Mice, Transgenic , Neutrophil Infiltration/immunology , Thyroid Neoplasms/immunology , Thyroid Neoplasms/pathology , Tumor BurdenABSTRACT
Tumor infiltrating lymphocytes have been correlated with a better prognosis for some tumors and medullary carcinoma of breast is a good example. However, in a recent study of invasive micropapillary carcinoma of breast, tumor infiltrating lymphocytes were associated with increased lymph node metastasis and a poorer prognosis. To explore possible mechanisms underlying this difference in immune responsiveness and tumor behavior, 28 cases of invasive micropapillary carcinoma with prominent lymphocyte infiltration were compared with 29 cases of medullary carcinoma. In both tumors, the majority of tumor infiltrating lymphocytes were T lymphocytes (P<0.01) with CD8+ T lymphocytes predominant (P<0.01). Significantly, functional differences in CD8+ cytotoxic T lymphocytes were identified in the two types of tumor. While lymphocytes infiltrated both the stroma and epithelial components of medullary carcinoma, the tumor infiltrating lymphocytes of invasive micropapillary carcinoma were almost exclusively confined to the stroma. Tumor infiltrating lymphocytes of medullary carcinoma showed stronger expression of FasL than those in invasive micropapillary carcinoma (P<0.01) and medullary carcinoma cells exhibited stronger expression of Fas than invasive micropapillary carcinoma cells did (P<0.01). In the subgroups of tumors with strong (++/+++) Fas expression, double immunohistochemistry revealed that most of the tumor infiltrating lymphocytes in medullary carcinoma, particularly those infiltrating the tumor nests, were CD8+ cytotoxic T lymphocytes, but not so in invasive micropapillary carcinoma. Furthermore, upregulated expression of perforin, granzyme B and FasL by cytotoxic T lymphocytes was greater in medullary carcinoma than invasive micropapillary carcinoma (P<0.01, respectively). The results suggest that effective immunity provided by tumor infiltrating lymphocytes varies in different tumors and the relative lack of tumor-killing cytotoxic T lymphocytes in invasive micropapillary carcinoma may explain, in part, the adverse association of tumor infiltrating lymphocytes with the biological behavior of invasive micropapillary carcinoma of breast.
Subject(s)
Breast Neoplasms/pathology , CD8-Positive T-Lymphocytes/pathology , Carcinoma, Medullary/pathology , Carcinoma, Papillary/pathology , Lymphocytes, Tumor-Infiltrating/pathology , T-Lymphocytes, Cytotoxic/pathology , Adult , Aged , Biomarkers, Tumor/metabolism , Breast Neoplasms/immunology , CD8-Positive T-Lymphocytes/immunology , Carcinoma, Medullary/immunology , Carcinoma, Papillary/immunology , Fas Ligand Protein/metabolism , Female , Humans , Immunoenzyme Techniques , Lymph Nodes/pathology , Lymphocytes, Tumor-Infiltrating/immunology , Middle Aged , Neoplasm Invasiveness , T-Lymphocytes, Cytotoxic/immunologyABSTRACT
CONTEXT: Patients with progressive medullary thyroid carcinoma (MTC) undergo multiple imaging procedures for diagnosis of relapse and staging. OBJECTIVE: Our objective was to assess the sensitivity and prognostic value of 18F-2-fluoro-2-deoxy-d-glucose (FDG) positron emission tomography (PET)/computed tomography (CT), and the imaging sensitivity of pretargeted iodine-131-radioimmunotherapy (RIT) in patients with progressive MTC. DESIGN/SETTING/PATIENTS: We performed a prospective multicenter study in high-risk patients with rapidly progressing MTC enrolled in a phase-II pretargeted RIT study, as documented by short serum calcitonin (Ct) or carcinoembryonic antigen (CEA) doubling time (DT). INTERVENTIONS/MAIN OUTCOME MEASURES: Patients underwent neck-thoracic-abdominal CT, spine and pelvic magnetic resonance imaging, whole-body post-RIT immunoscintigraphy (IS) with iodine-131, and whole-body 18F-FDG-PET/CT imaging. Imaging sensitivity and the correlation between FDG uptake and biomarkers DT were evaluated. RESULTS: A total of 33 patients with mean CEA and Ct DTs of 1.90 yr (range 0.21-8.50) and 1.52 yr (range 0.09-6.01), respectively, were evaluated. Sensitivity of FDG-PET/CT was 83% for neck, 85% for mediastinal, 75% for lung, 60% for liver, and 67% for bone metastases; overall sensitivity was 76%. Median standardized uptake value (SUVmax) was 5.23 (2.06-13.90). SUVmax correlated significantly with Ct DT (P = 0.011) and minimal DT (minimal value between CEA DT and Ct DT) (P = 0.027). Overall sensitivity of post-RIT IS, CT, and bone magnetic resonance imaging were 94, 74, and 85%, respectively. CONCLUSIONS: These results demonstrate the value of FDG-PET/CT for staging of patients with progressive MTC, especially in the neck and mediastinum, with possible prognostication by SUV quantification. Post-RIT IS was the most sensitive of the imaging modalities studied prospectively.
Subject(s)
Carcinoembryonic Antigen/immunology , Carcinoma, Medullary/diagnostic imaging , Carcinoma, Medullary/immunology , Fluorodeoxyglucose F18 , Radiopharmaceuticals , Thyroid Neoplasms/diagnostic imaging , Thyroid Neoplasms/immunology , Adult , Aged , Aged, 80 and over , Biomarkers/metabolism , Bone and Bones/diagnostic imaging , Female , Humans , Image Processing, Computer-Assisted , Iodine Radioisotopes , Magnetic Resonance Imaging , Male , Middle Aged , Prognosis , Prospective Studies , Radioimmunodetection , Radioimmunotherapy , Tomography, Emission-ComputedABSTRACT
Medullary thyroid carcinoma (MTC) is a relatively rare thyroid malignancy of C-cell origin that secretes calcitonin. Although its varied cytomorphologic features are well described in literature, very little is mentioned about the morphologic manifestation of its secretory activity. This study, based on nine fine needle aspiration (FNA) samples from eight MTC patients, is an attempt to present the varied cytomorphologic features suggesting secretory activity in MTC as observed in Papanicolaou and MGG stained FNA smears and correlate them with the immunocytochemical (ICC) staining for calcitonin performed on FNA smears and the serum calcitonin values. The average number of cells in these nine samples was as follows: oval/triangular/plasmacytoid (56.7%), small round (23.6%), spindle-shaped (12.7%), and miscellaneous (7.1%). The cytomorphological features suggesting secretory activity, viz., fine cytoplasmic vacuoles, azurophillic granules, marginal vacuoles, and intracytoplasmic lumina (ICL) with secretions were present in eight, eight, five, and six samples, respectively. Material likely to be amyloid, based on morphological features, was present extracellularly in three samples and both intracellularly and extracellularly in six samples. Immunocytochemically, all the nine samples stained for calcitonin and all the three stained for chromogranin showed positive cytoplasmic reaction in the neoplstic cells. The background amyloid (in six samples), the coarse cytoplasmic granules (in two samples), and the contents of ICL (in one sample) were found to be positively stained for calcitonin. The intracytoplasmic secretory material appeared to be diffusing out of some cells both in the routine MGG stained smears and in the smears stained for calcitonin. Histopathology reports of seven samples in six patients confirmed the cytodiagnosis of MTC in all. Baseline serum calcitonin values in three cases and postoperative serum calcitonin levels during follow-up in three others were high. Thus, our study highlighted the morphological manifestations of secretory activity in MTC and the nature of secretory material as calcitonin, supported by immunocytochemical staining and serum calcitonin level.
Subject(s)
Carcinoma, Medullary/metabolism , Thyroid Neoplasms/metabolism , Adult , Aged , Carcinoma, Medullary/immunology , Carcinoma, Medullary/pathology , Female , Humans , Immunohistochemistry , Male , Middle Aged , Thyroid Neoplasms/immunology , Thyroid Neoplasms/pathologyABSTRACT
BACKGROUND: Renal medullary carcinoma (RMC) is a rare kidney tumor that occurs in adolescent and young adults, typically in association with sickle cell trait. RMC exhibits rapid disease progression, frequent metastases at diagnosis, and dismal clinical outcomes. Currently available therapies, including cisplatin-based combination chemotherapy, multi-tyrosine kinase, and mTOR inhibitor strategies demonstrate either transient responses or minimal activity. Therefore, further molecular characterization and additional treatment strategies are urgently needed in this aggressive disease. The role of immune system surveillance and responsiveness to anti-PD-1 therapies in RMC are completely unexplored. CASE PRESENTATION: A 29 year old male with sickle cell trait presented with painless hematuria that ultimately resulted in a diagnosis of RMC. He underwent total nephrectomy and adjuvant cytotoxic chemotherapy with carboplatin, gemcitabine, paclitaxel, and bevacizumab. As is common in this aggressive form of kidney cancer he recurred with biopsy proven lymph node metastasis. He was started on checkpoint inhibitor therapy with nivolumab that inhibits program cell death protein 1 (PD-1), and on his first follow-up imaging he was found to have a partial response that on subsequent scans ultimately resulted in a complete response lasting greater than nine months. In this report, we present a patient with metastatic RMC who exhibited a clinical response to nivolumab, as well as the genetic and immunologic correlates of the pre-treatment tumor. Provocatively, robust immune infiltrate and expression of immune checkpoints were observed, despite the presence of a low mutation burden. CONCLUSIONS: Here, we report the first case of immune microenvironment profiling and response to anti-PD-1 in a patient with RMC to our knowledge. This case suggests that anti-PD-1 based therapies may have clinical activity in RMC.
Subject(s)
Carcinoma, Medullary/therapy , Kidney Neoplasms/therapy , Programmed Cell Death 1 Receptor/immunology , Tumor Microenvironment/drug effects , Adult , Antibodies, Monoclonal/administration & dosage , Carcinoma, Medullary/genetics , Carcinoma, Medullary/immunology , Carcinoma, Medullary/pathology , Female , Humans , Kidney Neoplasms/genetics , Kidney Neoplasms/immunology , Male , Neoplasm Metastasis , Nivolumab , Programmed Cell Death 1 Receptor/antagonists & inhibitors , Programmed Cell Death 1 Receptor/genetics , Tumor Microenvironment/immunologyABSTRACT
Invasive ductal carcinomas of breast with marked stromal lymphocytic infiltration have come to be classified as lymphocyte-predominant breast cancer (LPBC) because it obtains high pathological complete response rates with neoadjuvant chemotherapy. Medullary carcinoma (MC), which is independent from LPBC, is a rare histological subtype of invasive breast carcinoma accompanied by abundant lymphoplasmacytic infiltration as LPBC. Although MC shows marked cellular and structural atypia, it usually has a favorable outcome. It is occasionally difficult to distinguish MC from LPBC because both subtypes have nonspecific morphological features according to the present diagnostic criteria. Herein, we adopted multiplexed fluorescent immunohistochemistry to perform quantitative and simultaneous analyses of tumor-infiltrating lymphocytes (TILs) considering their spatial distribution and examined focal immune reaction differences between MC and LPBC. We found that CD8+ TILs are predominant in the intratumoral region, whereas CD4+ TILs are less common in MC. In non-luminal-type cancers, the numbers of stromal and intratumoral CD8+ TILs were significantly higher in MC than in LPBC. Stratified analyses by CD4+ TIL subsets showed robust infiltration of intratumoral CD8+ TILs in non-luminal-type MC even in suppressive environments, such a low T helper 1-to-regulatory T cell ratio. Our results suggest that extensive intratumoral CD8+ TIL infiltration might well be a promising biomarker for distinguishing MC from LPBC, especially in non-luminal-type cancers. Intratumoral CD8+ TILs and nonluminal intrinsic subtypes may serve as diagnostic characteristics allowing reliable histological criteria to be established for reproducibly diagnosing MC.
Subject(s)
Breast Neoplasms/immunology , CD4-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/immunology , Carcinoma, Medullary/immunology , Lymphocytes, Tumor-Infiltrating/immunology , Adult , Aged , Aged, 80 and over , Biomarkers, Tumor/analysis , Breast Neoplasms/pathology , CD4-Positive T-Lymphocytes/pathology , CD8-Positive T-Lymphocytes/pathology , Carcinoma, Medullary/pathology , Diagnosis, Differential , Female , Fluorescent Antibody Technique , Humans , Lymphocytes, Tumor-Infiltrating/pathology , Middle Aged , Neoplasm Invasiveness , Neoplasm Staging , Predictive Value of Tests , Retrospective Studies , Tumor MicroenvironmentABSTRACT
BACKGROUND: In vitro and in vivo studies have shown that dendritic cells (DCs) can stimulate antitumor T cell responses against medullary thyroid carcinoma (MTC). However, despite promising results in selected cases, the clinical efficacy of DC immunotherapy in patients with MTC has been limited. Recently, it has been demonstrated in mice that heat shock enhances the capacity of bone-marrow-derived DCs to stimulate antigen-specific T cells. The aim of our investigations was to evaluate whether heat shock also increases the capacity of human monocyte-derived DCs to stimulate antitumor T cell responses against MTC tumor cells. METHODS: DCs from six patients with metastatic MTC were pulsed with tumor lysate derived from allogeneic MTC tumor cells and were heat shocked for 12 h at 40 C or kept at 37 C. Thereafter, the DCs were matured and cocultured with T cells. Finally, the cytotoxic activity of T cells against MTC tumor cells was measured in vitro. RESULTS: In all patient samples, cytotoxic T cell responses against MTC tumor cells could be induced. Notably, heat-shocked DCs were more potent stimulators of cytotoxic T cell responses than control DCs, with T cells stimulated with heat-shocked DCs displaying a significantly increased cytotoxic activity against MTC tumor cells as compared with T cells stimulated with control DCs. In none of the experiments was a cytotoxic T cell response against unrelated pancreatic tumor cells (PANC-1) observed, using both control and heat-shocked DCs. CONCLUSIONS: Our study shows that heat-shocking DCs may be a valuable strategy to increase the immunostimulatory capacity of DCs used for immunotherapy of MTC.
Subject(s)
Carcinoma, Medullary/immunology , Dendritic Cells/metabolism , Dendritic Cells/pathology , Hyperthermia, Induced/methods , T-Lymphocytes, Cytotoxic/metabolism , Thyroid Neoplasms/immunology , Adult , Aged , Antigens, Surface/metabolism , Carcinoma, Medullary/pathology , Carcinoma, Medullary/therapy , Cell Differentiation , Cell Extracts/immunology , Female , Genes, MHC Class I , HLA Antigens/metabolism , HSP70 Heat-Shock Proteins/metabolism , HSP90 Heat-Shock Proteins/metabolism , Heat-Shock Response , Humans , Immunotherapy/methods , In Vitro Techniques , Lymphocyte Activation/immunology , Male , Middle Aged , Thyroid Neoplasms/pathology , Thyroid Neoplasms/therapy , Toll-Like Receptor 2/metabolism , Toll-Like Receptor 4/metabolism , Tumor Cells, CulturedSubject(s)
Breast Neoplasms/pathology , Carcinoma, Ductal, Breast/secondary , Carcinoma, Medullary/secondary , T-Lymphocytes, Regulatory/pathology , Adult , Aged , Biomarkers, Tumor/metabolism , Breast Neoplasms/immunology , Breast Neoplasms/metabolism , Carcinoma, Ductal, Breast/immunology , Carcinoma, Ductal, Breast/metabolism , Carcinoma, Medullary/immunology , Carcinoma, Medullary/metabolism , Cell Count , Female , Humans , Middle Aged , Neoplasms, Multiple PrimaryABSTRACT
The targeting potential of 131I-labeled NP-4 and MN-14 anti-CEA (carcinoembryonic antigen) monoclonal antibodies (MAbs) was assessed in 19 patients with metastatic medullary thyroid cancer (MTC). Seventeen of these patients also entered pilot radioimmunotherapy studies with nonmyeloablative doses of 131I-anti-CEA MAbs. Tumor targeting was possible in all 19 patients, with an overall lesion sensitivity of 91%. Tumor dosimetry with 131I-MN-14 IgG or F(ab)2 was very favorable, with tumor doses of 14.3 +/- 8.3 cGy/mCi and tumor:red marrow dose ratios exceeding 3:1 for most lesions. Limited antitumor effects lasting up to 26+ months, based on physical exam, tumor markers, computed tomography, or a followup MAb scan, were seen in 5 of 11 assessable patients given relatively low doses of 131I-labeled anti-CEA MAbs. We conclude that anti-CEA MAbs are excellent agents for targeting metastatic MTC. The high tumor uptake of the 131I-anti-CEA antibodies and evidence of tumor response in some patients suggest that radioimmunotherapy with radioiodinated anti-CEA MAbs may be an effective treatment for MTC, particularly when the maximum tolerated dose is given alone or in combination with autologous red marrow or peripheral stem cell support.
Subject(s)
Antibodies, Monoclonal/therapeutic use , Carcinoma, Medullary/radiotherapy , Iodine Radioisotopes/therapeutic use , Radioimmunotherapy , Thyroid Neoplasms/radiotherapy , Animals , Carcinoembryonic Antigen/immunology , Carcinoma, Medullary/diagnostic imaging , Carcinoma, Medullary/immunology , Female , Humans , Immunoglobulin Fab Fragments/therapeutic use , Male , Mice , Pilot Projects , Radionuclide Imaging , Thyroid Neoplasms/diagnostic imaging , Thyroid Neoplasms/immunologyABSTRACT
A minority of breast cancers is characterized by lymphoplasmacytic infiltrates that have been correlated with improved patient survivals. The association of improved prognosis with plasmacytic infiltrates has been classically linked with the rare medullary carcinoma subtype but is also evident in the smaller infiltrated fraction of the more abundant nonmedullary (not otherwise specified) tumors. It is our hypothesis that these plasma cell (PC) infiltrates represent a host humoral response driven by one or more tumor-derived neoantigens. As the index study group, two primary medullary carcinoma tumors were examined. Immunophenotyping confirmed a large number of IgG PCs in contradistinction to normal breast, which typically contains a lesser number of mainly IgA isotypes. IgG heavy and light chains were expressed as combinatorial phage Fab libraries. VH and VL sequences showed a preponderance of clonal groups in both patients, as identified by germ-line gene usage and junctional mutation patterns. Panning of phage Fab libraries against purified antigens excluded Her2/neu and p53 as the eliciting antigen, and failure of clonal enrichment by cell panning suggested that the neoantigen was not membrane expressed or was expressed at low levels. Cognate, original VH+VL pairs were obtained by single cell PCR of tumor PCs, which showed overlap with the pooled IgG libraries. Tumor-derived IgG V genes exhibited mutational patterns that were consistent with antigenic selection and affinity maturation. Where examined, IgG1 was the predominant isotype, consistent with a T-dependent (i.e., protein) antigen. From these data, we infer that the breast tumor PC infiltrates of the medullary carcinoma subtype are compatible with an autogenic tumor neoantigen-driven humoral immune response.