Increased Levels of PD1 and Glycolysis in CD4+ T Cells Promote Lymph Node Metastasis in Oral Squamous Cell Carcinoma Patients

Background: Cervical lymph node metastasis is one of the poorest prognostic factors in oral squamous cell carcinoma (OSCC). Activated immune cells and cancer cells generally have metabolic similarities in tumor microenvironment. However, it is unknown whether abnormal glycolysis in T cells could facilitate metastatic lymph nodes in patients with OSCC. Methods: Flow cytometry and immunofluorescence staining were used to analyze the differences in CD4+ PD1+ T cells between metastatic and negative lymph nodes. RT-PCR was performed to detail the expression of immune checkpoints and glycolysis-related enzymes in metastatic and negative lymph nodes. Kruskal-Wallis, Mann-Whitney, or nonparametric paired tests (i.e., the Wilcoxon matched paired test) were used to analyze the non-parametric distribution of the samples. Results: The frequency of CD4+ T cells decreased in the metastatic lymph nodes (p = 0.0019). Immune checkpoints (PD1, PDL1, and CTLA4) of CD4+ T cells were detected in metastatic (LN+) and paired negative lymph nodes (LN-) of OSCC patients. The PD1 expression of LN+ increased markedly compared to that of LN- (p = 0.0205). Similarly, the PD1 of CD4+ T cells in LN+ increased significantly compared to that of LN-. Glycolysis-related enzyme levels in CD4+ T cells from LN+ were dramatically higher than those in LN-. Moreover, PD1 and Hk2 expressions in CD4+ T cells increased in metastatic lymph nodes of OSCC patients with prior surgical treatment compared to those without. Conclusions: These findings suggest that increased PD1 and glycolysis in CD4+ T cells may serve as pivotal regulators of OSCC metastatic lymph nodes, which are closely associated with elevated glycolysis.

A Study on the Correlation Between M2 Macrophages and Regulatory T Cells in the Progression of Colorectal Cancer

Background: M2 macrophages and regulatory T cells (Tregs) can promote tumors and development by inhibiting the anti-tumor immune response. This study investigated the number of CD163‐positive M2 macrophages and Foxp3-positive Tregs in the progression of colorectal cancer. It also investigated the correlation and of M2 macrophages and Tregs.Methods: Postoperative tissue specimens and clinical data were collected from 197 patients with colorectal cancer who underwent initial surgical treatment in The Second Ward of Colorectal Surgery of the First Affiliated Hospital of Jinzhou Medical University from March 2020 to December 2020. Use immunohistochemical methods to detect the expression levels of CD163 protein-labeled M2 macrophages and Foxp3 protein-labeled Tregs in colorectal cancer tissues, matched paracancer tissues and lymph node tissues. Analyze the correlation between CD163 and Foxp3 in cancer tissues and lymph node tissues, as well as the relationship between clinicopathological characteristics and preoperative tumor markers. Results: M2 macrophages and Tregs were significantly positively correlated in cancer and lymph node tissues, which significantly increased in cancer and metastatic lymph node tissues. Interestingly, M2 macrophages in non-metastatic lymph nodes also increased significantly in patients with metastatic lymph nodes. Tregs stage I+II is higher than stage III+IV in paraneoplastic tissues. In addition, both CD163 and Foxp3 were upregulated with increasing tumor TNM stage, depth of infiltration, lymphatic metastasis, and depth of infiltration, and both were positively correlated with CEA. Conclusion: M2 macrophages and Tregs are important indicators of colorectal cancer progression and lymph node metastasis. There is a certain correlation between the two types of cells. It is possible that M2 macrophages, together with suppressor cells Tregs, promote an immunosuppressive environment.

Long-Term Efficacy of Ethanol Ablation as Treatment of Metastatic Lymph Nodes from Papillary Thyroid Carcinoma

Context Ethanol ablation (EA) is considered an alternative to surgery for metastatic lymph nodes from papillary thyroid carcinoma (PTC) in selected patients. Objective The aim of this study was to evaluate the long-term efficacy and safety of this particular treatment. Design and setting Adult patients with PTC who had received EA in lymph node metastasis at a tertiary referral center, and were included in a published study from 2011, were invited to participate in this follow-up study. Methods Radiologic- and medical history were reviewed. Ultrasound examination of the neck was performed by radiologists, and clinical examination was performed by an endocrine surgeon. Response was reported according to predefined criteria for satisfactory EA-treatment. Adverse events associated with EA were evaluated. Cause of death was reported for deceased patients. Results From the 2011-study 51 of 63 patients were included. Forty-four patients were reexamined (67/109 lesions) and 7 patients were deceased. Median follow-up time from primary surgery was 14.5 years. Median follow-up from the latest performed EA in the 2011 study was 11.3 years. Local control was permanently achieved in most patients (80 %). Recurrence within an ablated node was registered in 13 metastases in 10 patients. Seven of these patients also had recurrent disease elsewhere in the neck. No major side effects were reported. Conclusion EA is a minimally invasive procedure with a low risk of complications. Our data suggest that EA is a safe and efficient treatment, providing excellent results for a large group of patients also in the long term.

A novel targeted multifunctional nanoplatform for visual chemo-hyperthermia synergy therapy on metastatic lymph nodes via lymphatic delivery

Background Distant metastasis to vital organs is the major contributor to breast cancer mortality, and regional lymph node metastasis is an important facilitator of distant metastasis and recurrence in this cancer. The early diagnosis and precise treatment of lymph node metastasis are crucial for staging and prognosis in breast cancer. Herein, we report a visualized precision medicine nanoplatform of metastatic lymph nodes for ultrasonic/photoacoustic (US/PA) dual modal imaging-guided in situ targeted hyperthermia-combined chemotherapy. Results Carbon nanoparticles (CNs), approved by the China Food and Drug Administration, were loaded with docetaxel and rationally combined with anti-hypoxia-inducible factor 1α antibody-modified poly (lactic-co-glycolic acid) (PLGA) nanoparticles to achieve the combination of passive targeting at the lymph nodes and intracellular targeting at HIF 1α factor. The accumulation and retention of nanoparticles in metastatic lymph nodes via lymphatic delivery were enhanced. Docetaxel could be effectively offloaded by CNs that have active carbon nanoparticles, and the PLGA membrane prevented drug leakage. The nanoparticles exhibited excellent photothermal performance with a photothermal conversion efficiency of 28.9%, killing tumor cells in metastatic lymph nodes through hyperthermia. In vitro and in vivo systematic evaluations revealed that hyperpyrexia triggered the rupture of nanoparticles caused by the phase transition of perfluorohexane, resulting in docetaxel release for achieving in situ hyperthermia-combined chemotherapy. Conclusions The laser-triggered highly efficient in situ chemotherapy nanosystem achieves targeted synergistic chemo-hyperthermia treatment of metastatic lymph nodes, and lymphatic delivery represents a strategy to avoid additional injury caused by drugs entering the blood circulation. Graphical Abstract

Extrahepatic metastases of hepatocellular carcinoma on 18F FDG PET CT

Background To determine locations, relative frequencies, imaging features, and pattern of distribution of extrahepatic metastasis from hepatocellular carcinoma (HCC) on 2-deoxy-2-[fluorine-18]fluoro-D-glucose (18F-FDG) PET CT. Methods FDG PET CT scans of 224 consecutive patients of HCC acquired between 2010 and 2018 were reviewed. Fifty-six patients detected with extrahepatic metastasis on FDG PET CT were retrospectively analyzed. Findings were correlated with prior/follow-up imaging studies, clinical findings, FNAC, or biopsy findings whenever available. Descriptive analysis of location, relative frequencies, imaging features, and pattern of distribution of extrahepatic metastasis was done. Results Commonest were metastatic pulmonary nodules (55.3% patients), most of them being well-defined solid lesions (53.5%) with bilateral involvement in 44.6% patients and lower lobes of lungs along with other lobes being more frequently involved (41.0% patients). While in 7.14% patients lung nodules were FDG avid, 23.2% patients had both FDG avid and non-avid pulmonary nodules. Second most common were regional metastatic lymph nodes in 44.65% of patients seen at aortocaval (25%), paraaortic (23.21%), portocaval (21.4%), and left gastric nodal (17.8% of patients) stations. Twenty-five percent of patients had FDG avid lymph nodes and 5.36% patients had both FDG avid and FDG non-avid lymph nodes. Distant metastatic lymph nodes were third most common in 39.2% of patients seen at paratracheal (2.5%), juxtaphrenic (8.9%), and mesenteric lymphnodal (7.1%) stations. Twenty-five percent of patients had FDG avid lymph nodes while 5.36% patients had both FDG avid and FDG non-avid lymph nodes. Skeletal involvement was seen in 32.1% of patients. Commonest sites are vertebrae (16.7%), pelvis (14.2%), and ribs (10.7% patients). Six out of 7 patients had unilateral adrenal gland involvement. Bilateral adrenal gland involvement was seen in 1 patient. FDG non-avid peritoneal/omental metastases was seen in 2 patients. Brain, spleen, and muscle metastatic lesions were seen in 1 patient each out of 56 patients (1.79%). Conclusions Lungs, regional and distant lymph nodes and skeleton are the most frequently involved sites of extrahepatic metastatic hepatocellular carcinoma. Adrenal glands, muscles, brain and peritoneum are also involved but to a lesser extent.

Histomorphology and Molecular Genetics in Different Intra-Tumoral Zones and Matched Metastatic Lymph Nodes of Colorectal Cancer with Heterogenous Mismatch Repair Status

Objective: To better understand the clinicopathological characteristics and molecular alterations in different intra-tumoral components of colorectal cancer (CRC) with heterogeneity of mismatch repair (MMR) protein expression and microsatellite instability (MSI) status.Methods: We identified 4 cases of CRC with heterogenous MMR protein expression and analyzed the histopathological features, MSI status and other molecular alterations in separately microdissected intra-tumoral zones and lymph node metastases by polymerase chain reaction (PCR) -based MSI testing, MLH1 promoter methylation and targeted next-generation sequencing (NGS).Results: Microsatellite instability-high (MSI-H) was identified in the MLH1/PMS2 deficient zones in Case 1-3, and in the MSH2/MSH6 deficient zone in Case 4, while MSS was in all the intra-tumoral zones and metastatic lymph nodes with proficient MMR (pMMR). Furthermore, heterogeneity of MLH1 promoter methylation and/or other common driving gene mutations of CRC, such as KRAS, PIK3CA and so on, was identified in all the 4 CRCs. In addition, 75% (3/4) of cases showed heterogeneity of histomorphology in intra-tumoral components and metastatic lymph nodes (Case 1, 2, 4), and all the corresponding metastatic lymph nodes were moderate differentiation with MSS/pMMR (Case 2, 3). Conclusions: The heterogeneous MSI status is highly correlated with histomorphological heterogeneity, which is also an important clue for the heterogeneity of drive gene mutations in CRC. These results suggest that it is essential to detect MMR protein expression and other gene mutations in metastases before treatment, especially for the CRCs with heterogenous MMR protein expression or histomorphology.

Polypeptide-GalNAc-Transferase-13 Shows Prognostic Impact in Breast Cancer

Breast cancer is a public health concern and is currently the fifth cause of mortality worldwide. Identification of different biological subtypes is essential for clinical management; therefore, the role of pathologists is essential and useful tools for immunohistochemistry diagnosis are needed. Polypeptide-GalNAc-transferases are emerging novel biomarkers related to cancer behavior and GalNAc-T13, correlated with aggressiveness in some tumors, is an interesting candidate. Few monoclonal antibodies reacting with native proteins, and not affected by fixation and paraffin embedding, have been reported. The aim of this work was to develop a useful monoclonal antibody anti-GalNAc-T13 and to assess its potential significance in breast cancer diagnosis. We evaluated 6 human breast cancer cell lines, 338 primary breast tumors and 48 metastatic lymph nodes and looked for clinical significance correlating GalNAc-T13 expression with patients’ clinical features and survival. We found high GalNAc-T13 expression in 43.8% of the cases and observed a significant higher expression in metastatic lymph nodes, correlating with worse overall survival. We hypothesized several possible molecular mechanisms and their implications. We conclude that GalNAc-T13 may be a novel biomarker in breast cancer, useful for routine pathological diagnosis. Elucidation of molecular mechanisms related to aggressiveness should contribute to understand the role of GalNAc-T13 in breast cancer biology.