Background and Aims
Intrahepatic cholangiocarcinoma (ICC) is a severe malignant tumor in which the standard therapies are mostly ineffective. The biological significance of the desmoplastic tumor ...microenvironment (TME) of ICC has been stressed but was insufficiently taken into account in the search for classifications of ICC adapted to clinical trial design. We investigated the heterogeneous tumor stroma composition and built a TME‐based classification of ICC tumors that detects potentially targetable ICC subtypes.
Approach and Results
We established the bulk gene expression profiles of 78 ICCs. Epithelial and stromal compartments of 23 ICCs were laser microdissected. We quantified 14 gene expression signatures of the TME and those of 3 functional indicators (liver activity, inflammation, immune resistance). The cell population abundances were quantified using the microenvironment cell population‐counter package and compared with immunohistochemistry. We performed an unsupervised TME‐based classification of 198 ICCs (training set) and 368 ICCs (validation set). We determined immune response and signaling features of the different immune subtypes by functional annotations. We showed that a set of 198 ICCs could be classified into 4 TME‐based subtypes related to distinct immune escape mechanisms and patient outcomes. The validity of these immune subtypes was confirmed over an independent set of 368 ICCs and by immunohistochemical analysis of 64 ICC tissue samples. About 45% of ICCs displayed an immune desert phenotype. The other subtypes differed in nature (lymphoid, myeloid, mesenchymal) and abundance of tumor‐infiltrating cells. The inflamed subtype (11%) presented a massive T lymphocyte infiltration, an activation of inflammatory and immune checkpoint pathways, and was associated with the longest patient survival.
Conclusion
We showed the existence of an inflamed ICC subtype, which is potentially treatable with checkpoint blockade immunotherapy.
Here we present an optimized workflow for global proteome and phosphoproteome analysis of tissues or cell lines that uses isobaric tags (TMT (tandem mass tags)-10) for multiplexed analysis and ...relative quantification, and provides 3× higher throughput than iTRAQ (isobaric tags for absolute and relative quantification)-4-based methods with high intra- and inter-laboratory reproducibility. The workflow was systematically characterized and benchmarked across three independent laboratories using two distinct breast cancer subtypes from patient-derived xenograft models to enable assessment of proteome and phosphoproteome depth and quantitative reproducibility. Each plex consisted of ten samples, each being 300 μg of peptide derived from <50 mg of wet-weight tissue. Of the 10,000 proteins quantified per sample, we could distinguish 7,700 human proteins derived from tumor cells and 3100 mouse proteins derived from the surrounding stroma and blood. The maximum deviation across replicates and laboratories was <7%, and the inter-laboratory correlation for TMT ratio-based comparison of the two breast cancer subtypes was r > 0.88. The maximum deviation for the phosphoproteome coverage was <24% across laboratories, with an average of >37,000 quantified phosphosites per sample and differential quantification correlations of r > 0.72. The full procedure, including sample processing and data generation, can be completed within 10 d for ten tissue samples, and 100 samples can be analyzed in ~4 months using a single LC-MS/MS instrument. The high quality, depth, and reproducibility of the data obtained both within and across laboratories should enable new biological insights to be obtained from mass spectrometry-based proteomics analyses of cells and tissues together with proteogenomic data integration.
Fibroblasts have exceptional phenotypic plasticity and capability to secrete vast amount of soluble factors, extracellular matrix components and extracellular vesicles. While in physiological ...conditions this makes fibroblasts master regulators of tissue homeostasis and healing of injured tissues, in solid tumors cancer associated fibroblasts (CAFs) co‐evolve with the disease, and alter the biochemical and physical structure of the tumor microenvironment, as well as the behavior of the surrounding stromal and cancer cells. Thus CAFs are fundamental regulators of tumor progression and influence response to therapeutic treatments. Increasing efforts are devoted to better understand the biology of CAFs to bring insights to develop complementary strategies to target this cell type in cancer. Here we highlight components of the tumor microenvironment that play key roles in cancer progression and invasion, and provide an extensive overview of past and emerging understanding of CAF biology as well as the contribution that MS‐based proteomics has made to this field.
Matrix metalloproteinase (MMP)-8 and MMP-9 play several roles in inflammation, including degradation of extracellular matrix (ECM) components and regulation of cytokine activity. To determine the ...roles of MMP-8 and MMP-9 in a neutrophil-dependent inflammatory response, we used a murine model of corneal inflammation in which LPS is injected into the corneal stroma. In contrast to wild-type mice, we found that i) lipopolysaccharide (LPS)-injected CXCR2−/− corneas had impaired neutrophil infiltration and did not express either MMP-8 or MMP-9; ii) neutrophil migration through the central cornea was impaired in Mmp8−/− , but not Mmp9−/− , mice; iii) neutrophil migration was inhibited in collagenase-resistant mice; iv) the chemotactic Pro-Gly-Pro (PGP) tripeptide that binds CXCR2 was decreased in CXCR2−/− mice; v) PGP production was impaired in Mmp8−/− corneas; and vi) neutralizing anti-PGP antibody did not inhibit neutrophil infiltration in Mmp8−/− mice. We found no effects of MMP-8 on LPS-induced CXC chemokine (LIX, or CXCL5)-induced neutrophil recruitment or on LPS-induced CXC chemokine production. Together, these studies indicate that neutrophils contribute to the production of both MMP-8 and MMP-9 in LPS-injected corneas and that MMP-8 regulates neutrophil migration through the dense collagenous ECM of the corneal stroma by generating chemotactic PGP during inflammation.
Resistance of solid tumors to chemo‐ and radiotherapy remains a major obstacle in anti‐cancer treatment. Herein, the membrane protein caveolin‐1 (CAV1) came into focus as it is highly expressed in ...many tumors and high CAV1 levels were correlated with tumor progression, invasion and metastasis, and thus a worse clinical outcome. Increasing evidence further indicates that the heterogeneous tumor microenvironment, also known as the tumor stroma, contributes to therapy resistance resulting in poor clinical outcome. Again, CAV1 seems to play an important role in modulating tumor host interactions by promoting tumor growth, metastasis, therapy resistance and cell survival. However, the mechanisms driving stroma‐mediated tumor growth and radiation resistance remain to be clarified. Understanding these interactions and thus, targeting CAV1 may offer a novel strategy for preventing cancer therapy resistance and improving clinical outcomes. In this review, we will summarize the resistance‐promoting effects of CAV1 in tumors, and emphasize its role in the tumor‐stroma communication as well as the resulting malignant phenotype of epithelial tumors.
Micronodular thymoma with lymphoid stroma (MNT) is a rare thymic epithelial neoplasm subtype characterized by a micronodular tumor cell growth pattern and abundant lymphoid stroma. Micronodular ...thymic carcinoma with lymphoid stroma (MNCA) is considered as a malignant counterpart of MNT and exhibits a growth pattern similar to that of MNT but has histologic features reminiscent of thymic squamous cell carcinoma, such as cytologic atypia and CD5 and CD117 immunoexpression. Although both MNT and MNCA are characterized by abundant lymphoid stroma, it remains unknown whether there are differences in infiltrating lymphocytes between MNT and MNCA. We analyzed the immune microenvironment profile in eight MNT and three MNCA cases. The cell density of CD8‐positive T cells was significantly higher in MNT than in MNCA, whereas that of FOXP3‐positive T cells was significantly higher in MNCA than in MNT. There was no significant difference in the cell density of programmed death protein 1‐positive T cells and programmed death ligand 1 expression between the MNT and MNCA cases. Our findings indicated that the immune microenvironment of MNCA differed from that of MNT and, compared with the T‐cell profile of MNT, that of MNCA was more suppressive to patients′ antitumor immune response.
Hepatocellular carcinoma (HCC) is a primary liver cancer that usually develops in the setting of chronic inflammation and liver damage. The hepatic microenvironment plays a crucial role in the ...disease development, as players such as hepatic stellate cells, resident liver macrophages (Kupffer cells), endothelial cells, extracellular matrix, and a variety of immune cells interact in highly complex and intertwined signaling pathways. A key factor in these cross-talks are platelets, whose role in cancer has gained growing evidence in recent years. Platelets have been reported to promote HCC cell proliferation and invasion, but their involvement goes beyond the direct effect on tumor cells, as they are known to play a role in pro-fibrinogenic signaling and the hepatic immune response, as well as in mediating interactions between these factors in the stroma. Anti-platelet therapy has been shown to ameliorate liver injury and improve the disease outcome. However, platelets have also been shown to play a crucial role in liver regeneration after organ damage. Therefore, the timing and microenvironmental setting need to be kept in mind when assessing the potential effect and therapeutic value of platelets in the disease progression, while further studies are needed for understanding the role of platelets in patients with HCC.
To evaluate and compare the depth of corneal stromal demarcation line after accelerated collagen crosslinking (CXL) using continuous and pulsed light ultraviolet-A (UVA) exposure.
Department of ...Ophthalmology, Arcispedale Santa Maria Nuova, Reggio Emilia, Italy.
Retrospective case series.
Patients with progressive keratoconus were assigned to 1 of 2 treatment protocols using the same irradiation device for accelerated CXL. Patients assigned to Group A received accelerated CXL using continuous UVA light exposure at 30 mW/cm(2) for 4 minutes. Patients assigned to Group B received accelerated CXL using pulsed UVA light with 8 minutes (1 second on/1 second off) of UVA exposure at 30 mW/cm(2) and energy dose of 7.2 J/cm(2). One month after surgery, corneal stromal demarcation line depth was measured by 2 independent observers using anterior segment optical coherence tomography (AS-OCT).
A total of 60 patients were assessed. Corneal stromal demarcation line was easily identified on AS-OCT scans in all eyes by both observers. The mean depth of stromal demarcation line was 149.32 ± 36.03 μm in Group A and 213 ± 47.38 μm in Group B. The difference in stromal demarcation line depth between groups was statistically significant (P < .001).
Using accelerated CXL, the corneal stromal demarcation line was significantly deeper using pulsed rather than continuous light exposure.
No author has financial or proprietary interest in any material or method mentioned.
To evaluate the clinical outcomes of a new model of intrastromal corneal ring segments (ICRSs) (Keraring AS) in patients with keratoconus and quantify subsequent changes in refraction and corneal ...topography.
This nonrandomized, single-center, retrospective observational study explores the effect of progressive thickness ICRS implantation in patients with keratoconus with a 3-month follow-up. After creating an intrastromal tunnel using a femtosecond laser, 1 or 2 ICRSs of progressive thicknesses (150/250 μm or 200/300 μm) and 160-degree arc length were implanted. Changes in uncorrected distance visual acuity, best-corrected distance visual acuity, refractive outcomes, corneal astigmatism, and maximum keratometry readings were recorded before and after surgery.
The study cohort consisted of 82 patients (104 eyes) with a mean age of 31.2 ± 10 years. At 3 months, ICRS implantation significantly improved uncorrected and corrected visual acuities from 0.82 to 0.46 (logarithm of the minimum angle of resolution LogMAR) and from 0.31 to 0.21 (LogMAR), respectively (P < 0.001). The mean spherical error reduced from -1.74 diopters (D) to -0.90 D (P < 0.001), and the mean cylindrical error reduced from -4.22 D to -2.01 D (P < 0.001). The manifest refraction spherical equivalent reduced from -3.85 D to -1.91 D (P < 0.001). All topographic parameters were reduced, including maximum keratometry (53.6 D vs. 50.3 D) and keratometric astigmatism (-4.6 D vs. -2.2 D) (P < 0.001).
The Keraring AS provides a new, apparently safe, and effective means of improving visual acuity and reducing the refractive error and mean keratometry in eyes with asymmetric keratoconus.
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