Intra-tumor heterogeneity is one of the biggest challenges in cancer treatment today. Here we investigate tissue-wide gene expression heterogeneity throughout a multifocal prostate cancer using the ...spatial transcriptomics (ST) technology. Utilizing a novel approach for deconvolution, we analyze the transcriptomes of nearly 6750 tissue regions and extract distinct expression profiles for the different tissue components, such as stroma, normal and PIN glands, immune cells and cancer. We distinguish healthy and diseased areas and thereby provide insight into gene expression changes during the progression of prostate cancer. Compared to pathologist annotations, we delineate the extent of cancer foci more accurately, interestingly without link to histological changes. We identify gene expression gradients in stroma adjacent to tumor regions that allow for re-stratification of the tumor microenvironment. The establishment of these profiles is the first step towards an unbiased view of prostate cancer and can serve as a dictionary for future studies.
To describe the surgical technique of performing a wedge resection of a 1 mm area of inferior corneal stroma using stromal air injection assisted separation sparring the endothelium in a patient with ...bilateral Pellucid Marginal Degeneration. 68-year-old male Caucasian advanced, non progressive. A 30G needle on a 1 ml air-filled syringe injects air into the stromal lamellae with the bevel up, leading to stromal emphysema. An MVR blade is used to incise the marked area until clear corneal tissue is seen. The incised areas are separated with a lamellar dissector, and the crescentic area is excised. The lips of the wound are approximated with 10-0 Prolene sutures. This surgical approach leads to adequate management of the Pellucid Marginal Degeneration without injury to the endothelium, ensuring reduced antigenic input and maintenance of the architectural integrity of the eye.
Although the tumor-stroma ratio (TSR) has prognostic value in many cancers, the traditional semi-quantitative visual assessment method has inter-observer variability, making it impossible for ...clinical practice. We aimed to develop a machine learning (ML) algorithm for accurately quantifying TSR in hematoxylin-and-eosin (H&E)-stained whole slide images (WSI) and further investigate its prognostic effect in patients with muscle-invasive bladder cancer (MIBC). We used an optimal cell classifier previously built based on QuPath open-source software and ML algorithm for quantitative calculation of TSR. We retrospectively analyzed data from two independent cohorts to verify the prognostic significance of ML-based TSR in MIBC patients. WSIs from 133 MIBC patients were used as the discovery set to identify the optimal association of TSR with patient survival outcomes. Furthermore, we performed validation in an independent external cohort consisting of 261 MIBC patients. We demonstrated a significant prognostic association of ML-based TSR with survival outcomes in MIBC patients (
< 0.001 for all comparisons), with higher TSR associated with better prognosis. Uni- and multivariate Cox regression analyses showed that TSR was independently associated with overall survival (
< 0.001 for all analyses) after adjusting for clinicopathological factors including age, gender, and pathologic stage. TSR was found to be a strong prognostic factor that was not redundant with the existing staging system in different subgroup analyses (
< 0.05 for all analyses). Finally, the expression of six genes (DACH1, DEEND2A, NOTCH4, DTWD1, TAF6L, and MARCHF5) were significantly associated with TSR, revealing possible potential biological relevance. In conclusion, we developed an ML algorithm based on WSIs of MIBC patients to accurately quantify TSR and demonstrated its prognostic validity for MIBC patients in two independent cohorts. This objective quantitative method allows application in clinical practice while reducing the workload of pathologists. Thus, it might be of significant aid in promoting precise pathology services in MIBC.
Disorders of the transparent cornea affect millions of people worldwide. However, how to maintain and/or regenerate this organ remains unclear. Here, we show that
(encoding a canonical NF-κB subunit) ...ablation in K14
corneal epithelial stem cells not only disrupts corneal regeneration but also results in age-dependent epithelial deterioration, which triggers aberrant wound-healing processes including stromal remodeling, neovascularization, epithelial metaplasia, and plaque formation at the central cornea. These anomalies are largely recapitulated in normal mice that age naturally. Mechanistically,
deletion suppresses expression of Aldh1a1, an enzyme required for retinoic acid synthesis from vitamin A. Retinoic acid administration blocks development of ocular anomalies in
mice and naturally aged mice. Moreover, epithelial metaplasia and plaque formation are preventable by inhibition of angiogenesis. This study thus uncovers the major mechanisms governing corneal maintenance, regeneration, and aging and identifies the NF-κB-retinoic acid pathway as a therapeutic target for corneal disorders.
Renal cell carcinoma with fibromyomatous stroma (RCC FMS), defined as an "emerging entity" in the 2016 WHO classification and recommended to be a novel entity by GUPS, is represented by tumor cells ...with clear to mildly eosinophilic cytoplasm displaying elongated and branching tubules and papillae. A fibromyomatous stroma could be observed in these tumors. These tumors are immunopositive for CK7 and featured by ELOC and/or TSC/mTOR gene mutations. In the 2022 WHO classification, ELOC mutated RCC is classified as a molecularly defined RCCs as an individual renal entity. However, there are limited descriptions of TSC/mTOR alterations in RCC FMS. Herein, we reported a case of 28-year-old woman with RCC FMS with intact ELOC and TSC/mTOR genes but ASXL1 mutation. The tumor cells were positive for mTOR expression. This case may indicate that altered mTOR expression, but not limited to mutated TSC/mTOR gene, that participates in the pathogenesis of RCC FMS.
Purpose
In biliary tract cancer (BTC), malignancy is strongest at the invasion front. To improve the BTC prognosis, the invasion front should be controlled. We evaluated tumor–stroma crosstalk at the ...tumor center and at the invasion front of BTC lesions. We investigated the expression of SPARC, a marker of cancer-associated fibroblasts, and determined its ability to predict BTC prognosis after neoadjuvant chemoradiotherapy (NAC-RT).
Methods
We performed immunohistochemistry to evaluate SPARC expression in resected specimens from patients that underwent BTC surgery. We established highly invasive (HI) clones in two BTC cell lines (NOZ, CCLP1), and performed mRNA microarrays to compare gene expression in parental and HI cells.
Results
Among 92 specimens, stromal SPARC expression was higher at the invasion front than at the lesion center (
p
= 0.014). Among 50 specimens from patients treated with surgery alone, high stromal SPARC expression at the invasion front was associated with a poor prognosis (recurrence-free survival:
p
= 0.033; overall survival:
p
= 0.017). Coculturing fibroblasts with NOZ-HI cells upregulated fibroblast SPARC expression. mRNA microarrays showed that connective tissue growth factor (CTGF) was upregulated in NOZ-HI and CCLP1-HI cells. A CTGF knockdown suppressed cell invasion in NOZ-HI cells. Exogeneous CTGF upregulated SPARC expression in fibroblasts. SPARC expression at the invasion front was significantly lower after NAC-RT, compared to surgery alone (
p
= 0.003).
Conclusion
CTGF was associated with tumor–stroma crosstalk in BTC. CTGF activated stromal SPARC expression, which promoted tumor progression, particularly at the invasion front. SPARC expression at the invasion front after NAC-RT may serve as a prognosis predictor.
The tumour-stroma ratio (TSR) and tumour budding (TB) are two high-risk factors with potential to be implemented in the next TNM classification. The aim of the current study was to evaluate the ...practical application of the two biomarkers based on reproducibility, independency and prognostic value. Patients diagnosed with stage II or III colon cancer who underwent surgery between 2005 and 2016 were included. Both TSR and TB were scored on haematoxylin and eosin-stained tissue sections. The TSR, based on the relative amount of stroma, was scored in increments of 10%. TB was scored following the consensus guidelines; a bud was defined as ≤ 4 tumour cells. For analysis, three categories were used. Cohen’s kappa was used for reproducibility. The prognostic value was determined with survival analysis. In total, 246 patients were included. The TSR distribution was
N
= 137 (56%) stroma-low and
N
= 109 (44%) stroma-high. The TB distribution was TB-low
N
= 194 (79%), TB-intermediate
N
= 35 (14%) and TB-high
N
= 17 (7%). The reproducibility of the TSR was good (interobserver agreement kappa = 0.83 and intraobserver agreement kappa = 0.82), whereas the inter- and intraobserver agreement for scoring TB was moderate (kappa 0.47 and 0.45, respectively). The survival analysis showed an independent prognostic value for disease-free survival for TSR (HR 1.57; 95% CI 1.01–2.44;
p
= 0.048) and for TB-high (HR 2.01; 95% CI 1.02–3.96;
p
= 0.043). Based on current results, we suggest the TSR is a more reliable parameter in daily practice due to better reproducibility and independent prognostic value for disease-free survival.
Antibody drugs have become the mainstream of cancer treatment due to advances in cancer biology and Ab engineering. However, several barriers to Ab therapy have also been identified. These include ...various mechanisms for Ab drug resistance, such as heterogeneity of antigen expression in tumor cells and reduction in antitumor immunity due to expression diversity, polymorphism of Fc receptors (FcR) in effector cells, and reduced function of effector cells. Countermeasures to each resistance mechanism are being investigated. This review focuses on barriers that impede the delivery of Ab drugs due to features of the solid tumor microenvironment. Unlike hematological malignancies, in which the target tumor cells are in blood vessels, clinical solid tumors contain cancer stroma, which interferes with the delivery of Ab drugs. In addition, the cancer mass itself interferes with the penetration of Ab drugs. In this article, I will consider the etiology of cancer stroma and propose a new Ab drug development strategy for solid cancer treatment centering on cancer stromal targeting (CAST) therapy using anti‐insoluble fibrin Ab‐drug conjugate (ADC), which can overcome the cancer stroma barrier. The recent success of ADCs, chimeric antigen receptor T cells (CAR‐Ts), and Bi‐specific Abs is changing the category of Ab drugs from molecular‐targeted drugs based on growth signal inhibition to cancer‐specific targeted therapies. Therefore, at the end of this review, I argue that it is time to reorient the concept of Ab drug development.
This review focuses on the lesser‐known barrier that impedes the delivery of Ab drugs due to features of the solid tumor microenvironment. With the recent success of chimeric antigen receptor T cell (CAR‐T) therapy and anti‐insoluble fibrin Ab‐drug conjugate (ADC), Ab development strategies have changed significantly, largely because CAR‐T and ADC do not need to neutralize growth signals. Instead, cancer specificity is essential in the new tactics for Ab therapeutics.
To evaluate the initial outcomes of femtosecond intrastromal lenticular implantation (FILI) combined with accelerated collagen cross-linking in patients with progressive keratoconus.
In this ...interventional, prospective, exploratory case series, patients with progressive keratoconus and contact lens intolerance were included. All eyes underwent femtosecond laser-enabled placement of stromal donor tissue and simultaneous accelerated collagen cross-linking. Follow-up of patients was conducted for a mean period of 190 ± 13 days (range, 177-193 days).
Six eyes from 6 patients were included in the study. Based on values before and 6 months after the procedure, clinical improvement was noted in uncorrected distance visual acuity (1.06 ± 0.48 logMAR vs. 0.38 ± 0.27 logMAR), corrected distance visual acuity (0.51 ± 0.20 logMAR vs. 0.20 ± 0.24 logMAR), and manifest spherical equivalent (-3.47 ± 1.15 D vs. -1.77 ± 1.7 D). There was flattening of mean keratometry in 3-mm and 5-mm zones by 3.42 ± 2.09 D and 1.70 ± 1.31 D, respectively. Mean pachymetry in the central and midperipheral zones increased by 18.3 ± 7.3 μm and 33.0 ± 8.8 μm, respectively. All eyes had reduction in higher-order aberrations, specifically coma. No eye lost lines of corrected distance visual acuity. No adverse events such as haze, infection, or allogeneic graft rejection were observed.
Initial experience with this small number of eyes suggests that the combination of tissue addition and accelerated collagen cross-linking may be a feasible option for low to moderate keratoconus. A larger cohort and longer follow-up are required to validate our results and establish long-term safety and efficacy of the procedure.
Maturation of the endothelium and the adjacent matrix was characterized in wild-type (WT) mice. The influence of FACIT collagen XII and XIV deficiency on the morphology, maturation, and function of ...the corneal endothelium was examined.
Analysis of the endothelium and Descemet's membrane (DM) was performed using transmission electron microscopy at postnatal day (P)4, P14, and P30 in WT, Col12a1(-/-), Col14a1(-/-), and Col12a1(-/-)/Col14a1(-/-) mice. Endothelial junctions were analyzed using ZO-1. The presence of endothelial-stromal communications was evaluated with phalloidin staining as well as electron microscopy. Finally, corneal thickness was assessed.
A thin DM, clefts between endothelial cells and DM, and large "vacuole-like" structures were present in the endothelial cells of WT mice at P4 but not noted at P30. The endothelia of Col12a1(-/-), Col14a1(-/-), and compound Col12a1(-/-)/Col14a1(-/-) in the P30 cornea maintained the vacuole-like structures seen at P4. A mature endothelial junction pattern was delayed in the null corneas. Expression of ZO-1 in WT endothelia at P14 was diffuse and localized to the basolateral and apical cell membrane. At P30, staining was localized to intercellular junctions. ZO-1 reactivity was patchy in Col12a1(-/-), Col14a1(-/-), and compound Col12a1(-/-)/Col14a1(-/-) corneas at P14 and P30. Stromal thickness was increased in P30 null corneas. Endothelial cell processes were demonstrated penetrating the DM and into the underlying stroma, throughout the entire endothelial layer in the P4 cornea.
Collagen XII and XIV null mice demonstrate delayed endothelial maturation. The structural alterations suggest functional changes in endothelial function resulting in increased corneal thickness. Endothelial-stromal interactions suggest a pathway for signal transduction.
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