To examine the effect of riboflavin/UVA corneal crosslinking on stromal ultrastructure and hydrodynamic behaviour.
One hundred and seventeen enucleated ungulate eyes (112 pig and 5 sheep) and 3 pairs ...of rabbit eyes, with corneal epithelium removed, were divided into four treatment groups: Group 1 (28 pig, 2 sheep and 3 rabbits) were untreated; Group 2 (24 pig) were exposed to UVA light (3.04 mW/cm(2)) for 30 minutes and Group 3 (29 pig) and Group 4 (31 pig, 3 sheep and 3 rabbits) had riboflavin eye drops applied to the corneal surface every 5 minutes for 35 minutes. Five minutes after the initial riboflavin instillation, the corneas in Group 4 experienced a 30 minute exposure to UVA light (3.04 mW/cm(2)). X-ray scattering was used to obtain measurements of collagen interfibrillar spacing, spatial order, fibril diameter, D-periodicity and intermolecular spacing throughout the whole tissue thickness and as a function of tissue depth in the treated and untreated corneas. The effect of each treatment on the hydrodynamic behaviour of the cornea (its ability to swell in saline solution) and its resistance to enzymatic digestion were assessed using in vitro laboratory techniques.
Corneal thickness decreased significantly following riboflavin application (p<0.01) and also to a lesser extent after UVA exposure (p<0.05). With the exception of the spatial order factor, which was higher in Group 4 than Group 1 (p<0.01), all other measured collagen parameters were unaltered by cross-linking, even within the most anterior 300 microns of the cornea. The cross-linking treatment had no effect on the hydrodynamic behaviour of the cornea but did cause a significant increase in its resistance to enzymatic digestion.
It seems likely that cross-links formed during riboflavin/UVA therapy occur predominantly at the collagen fibril surface and in the protein network surrounding the collagen.
No other tissue in the body depends more on the composition and organization of the extracellular matrix (ECM) for normal structure and function than the corneal stroma. The precise arrangement and ...orientation of collagen fibrils, lamellae and keratocytes that occurs during development and is needed in adults to maintain stromal function is dependent on the regulated interaction of multiple ECM components that contribute to attain the unique properties of the cornea: transparency, shape, mechanical strength, and avascularity. This review summarizes the contribution of different ECM components, their structure, regulation and function in modulating the properties of the corneal stroma. Fibril forming collagens (I, III, V), fibril associated collagens with interrupted triple helices (XII and XIV), network forming collagens (IV, VI and VIII) as well as small leucine-rich proteoglycans (SLRP) expressed in the stroma: decorin, biglycan, lumican, keratocan, and fibromodulin are some of the ECM components reviewed in this manuscript. There are spatial and temporal differences in the expression of these ECM components, as well as interactions among them that contribute to stromal function. Unique regions within the stroma like Bowman's layer and Descemet's layer are discussed. To define the complexity of corneal stroma composition and structure as well as the relationship to function is a daunting task. Our knowledge is expanding, and we expect that this review provides a comprehensive overview of current knowledge, definition of gaps and suggests future research directions.
•The unique corneal stromal structure and function is regulated by different extracellular matrix components.•The stroma proper as well as Bowman’ and Descemet’slayers have unique ECM compositions and functions.•Regulated collagen fibril formation is critical for stromal structure and function.•ECM molecules including SLRPs regulate stromal fibrillogenesis, hydration, and the availability of growth factors.•Regulation of ECM components play a major role in wound healing.
Objective
Tumor‐stroma ratio (TSR) is a promising parameter representing the abundance of the stroma which has been validated in many solid tumors. However, it is still not clear which part of stroma ...mainly contribute to the prognostic value of TSR. The aim of this study is to confirm the prognostic value of TSR in a large cohort of oral squamous cell carcinoma (OSCC) and further demonstrated that cancer‐associated fibroblasts (CAFs)‐stroma ratio (CSR) contributed to the prognostic value of TSR.
Materials and methods
TSR was evaluated on hematoxylin and eosin stained tissue samples from 581 patients with OSCC, which divides patients into high (>50%) and low (<50%) stroma. Then, CSR was estimated on immunohistochemical staining slides of 100 patients selected from 581 patients.
Results
In multivariate analysis, TSR was identified as an independent prognostic factor for disease‐free survival (DFS) (p < 0.001) and oral cancer‐specific survival (OCSS) (p < 0.001). The interaction term reached statistical significance for histological grade for DFS and OCSS separately. Furthermore, the high‐stroma group had a higher CSR than the low‐stroma group.
Conclusion
The prognostic value of TSR is validated in OSCC particularly in moderate and high differentiation, and CSR plays its part in the prognosis of TSR.
There are multiple challenges facing the current T cell therapies. First of all, the current CAR-T or TCR-T cell therapy targets single/dual antigens on tumor cells to kill tumor cells, which ...requires the infused CAR-T cells must survive a long duration of time without exhausting to eradicate tumors. Even if the survival challenge is resolved, the next challenge for eliminating solid tumors is the heterogenity, meaning some tumor cells that do not bear the targeted antigens will esxape from the treatment. Moreover, regardless CAR-T, TILs or TCR-T cells have to face the another major challenge of penetrating the stroma rich tumors and avoiding to be trapped in the stroma in order to kill tumor cells. Finally, some CAR-T cells may cause severe peripheral tissue damage associated toxicity. In this presentation, we aims to offer a totally different approach to address the listed challenges facing T cell therapy.
After two decades of effort from my group, we have invented an attIL12-T cell therapy and launched a clinical trial to address the listed challenges described in the background. This attIL12-T cell therapy is based on our multiple key discoveries during the past 2 decades from us: 1) translocation of vimentin from intracellular to cell surface in the highly malignant tumor cells across types, referred to as cell surface vimentin (CSV); 2) revealation of the CSV binding mini-peptide ligand VNTANST; 3) linking this VNTANST peptide to IL12 via genetic engineering yields a tumor-targeted IL12 (ttIL12) that reduces Fox3 Treg and MDSC in metastatic tumors, inducing memory T cells to reject tumor cell rechallenge; and 4) anchoring this ttIL12 on the surface of T cells (referred to as attIL12-T cells) eliminates heterogenous PDX osteosarcomas in immune deficient mice. In immune competent mice, this attIL12-OT1 T cells induces tumor antigen spreading against tumor cell rechallenge from tumor free mice. These achievements via this attIL12-T cell therapy is through destruction of CAFs in tumor stroma, boosting infused T cell accumulation in tumors and causing tumor cell death via multiple mechanism, which will be discussed in the meeting. A clinical trial has been launched this septement to validate these preclinical discoveries in human patient, which will also be discussed in this conference.
Regeneration of corneal stroma has always been a challenge due to its sophisticated structure and keratocyte-fibroblast transformation. In this study, we fabricate grid poly (ε-caprolactone)-poly ...(ethylene glycol) microfibrous scaffold and infuse the scaffold with gelatin methacrylate (GelMA) hydrogel to obtain a 3 D fiber hydrogel construct; the fiber spacing is adjusted to fabricate optimal construct that simulates the stromal structure with properties most similar to the native cornea. The topological structure (3 D fiber hydrogel, 3 D GelMA hydrogel, and 2 D culture dish) and chemical factors (serum, ascorbic acid, insulin, and β-FGF) are examined to study their effects on the differentiation of limbal stromal stem cells to keratocytes or fibroblasts and the phenotype maintenance, in vitro and in vivo tissue regeneration. The results demonstrate that fiber hydrogel and serum-free media synergize to provide an optimal environment for the maintenance of keratocyte phenotype and the regeneration of damaged corneal stroma.
Liver cancer is the second leading cause of cancer mortality worldwide, causing more than 700,000 deaths annually. Because of the wide landscape of genomic alterations and limited therapeutic success ...of targeting tumor cells, a recent focus has been on better understanding and possibly targeting the microenvironment in which liver tumors develop. A unique feature of liver cancer is its close association with liver fibrosis. More than 80% of hepatocellular carcinomas (HCCs) develop in fibrotic or cirrhotic livers, suggesting an important role of liver fibrosis in the premalignant environment (PME) of the liver. Cholangiocarcinoma (CCA), in contrast, is characterized by a strong desmoplasia that typically occurs in response to the tumor, suggesting a key role of cancer-associated fibroblasts (CAFs) and fibrosis in its tumor microenvironment (TME). Here, we discuss the functional contributions of myofibroblasts, CAFs, and fibrosis to the development of HCC and CCA in the hepatic PME and TME, focusing on myofibroblast- and extracellular matrix-associated growth factors, fibrosis-associated immunosuppressive pathways, as well as mechanosensitive signaling cascades that are activated by increased tissue stiffness. Better understanding of the role of myofibroblasts in HCC and CCA development and progression may provide the basis to target these cells for tumor prevention or therapy.
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