Invasive Front Research Articles

The impact of tumor budding and single-cell invasion on survival in patients with stage III/IV locally advanced oral squamous cell carcinoma- results from a prospective cohort study.

Tumor budding (TB) refers to the presence of small clusters of tumor cells at the invasive front of a malignant tumor. Single tumor cell invasion (SCI) is an extreme variant of TB, in which individual loose tumor cells are present at the invasive front. Both TB and SCI are important histomorphologic risk factors postulated to indicate loss of cellular cohesion. In this study, we investigated the influence of TB and SCI on different survival outcomes in patients with locally advanced oral squamous cell carcinoma (OSCC). We included 129 patients with locally advanced OSCC (pT3-4) from a single-center, prospectively maintained cohort. We examined the association of TB and SCI with the presence of occult lymph node metastasis using a logistic regression model. Survival probabilities were estimated using the Kaplan-Meier method and cumulative incidence functions. The association of TB and SCI on overall survival (OS), oral cancer-specific survival (OCSS), and local recurrence-free survival (LRFS) was investigated using Cox's proportional hazards regression models. TB was detected in 98 (76%) of the tumors, while SCI was observed in 66 (51%) patients. There was a significant association between TB and the occurrence of occult lymph node metastasis (OR=3.33, CI: 1.21-10.0). On multivariate analysis, TB had no detectable impact on survival outcomes. However, SCI showed a higher risk for local recurrence (Hazards ratio (HR): 3.33, CI: 1.19 - 9.27). This study demonstrates that TB and SCI in locally advanced OSCC function as an independent risk factor for occult lymph node metastases, as well as local recurrences. Both histomorphologic risk factors could serve as an additional parameter for stratifying therapy and escalating multimodal treatment approaches.

A Tissue Engineered 3D Model of Cancer Cell Invasion for Human Head and Neck Squamous-Cell Carcinoma.

Head and neck squamous-cell carcinoma (HNSCC) is associated with aggressive local invasiveness, being a main reason for its poor prognosis. The exact mechanisms underlying the strong invasive abilities of HNSCC remain to be elucidated. Therefore, there is a need for in vitro models to study the interplay between cancer cells and normal adjacent tissue at the invasive tumor front. To generate oral mucosa tissue models (OMM), primary keratinocytes and fibroblasts from human oral mucosa were isolated and seeded onto a biological scaffold derived from porcine small intestinal submucosa with preserved mucosa. Thereafter, we tested different methods (single tumor cells, tumor cell spots, spheroids) to integrate the human cancer cell line FaDu to generate an invasive three-dimensional model of HNSCC. All models were subjected to morphological analysis by histology and immunohistochemistry. We successfully built OMM tissue models with high in vivo-in vitro correlation. The integration of FaDu cell spots and spheroids into the OMM failed. However, with the integration of single FaDu cells into the OMM, invasive tumor cell clusters developed. Between segments of regular epithelial differentiation of the OMM, these clusters showed a basal membrane penetration and lamina propria infiltration. Primary human fibroblasts and keratinocytes seeded onto a porcine carrier structure are suitable to build an OMM. The HNSCC model with integrated FaDu cells could enable subsequent investigations into cancer cell invasiveness.

Lionfish (Pterois miles) in the Mediterranean Sea: a review of the available knowledge with an update on the invasion front

Invasive species often severely impact ecosystems and human activities in the areas that they invade. The lionfishes Pterois miles and P. volitans are regarded as the most successful invasive fishes in marine ecosystems. In the last 40 years, these Indo-Pacific predators have established in the tropical western Atlantic Ocean, with well-documented detrimental effects on the local fish communities. Around 10 years ago, a second invasion began in the Mediterranean Sea, which is being colonised by P. miles. Given the invasive potential of P. miles and the fact that the ecology and biodiversity of the temperate/sub-tropical Mediterranean Sea offer a different setting from the tropical western Atlantic, specific knowledge on this second invasion is needed. Here, we: (i) review the scientific knowledge available on the ecology of invasive lionfishes, (ii) discuss such knowledge in the context of invasion ecology and (iii) suggest future research avenues on the P. miles invasion in the Mediterranean Sea. In addition, we offer an update on the spread of P. miles in the Mediterranean Sea. While the history and development of the Mediterranean invasion are resolved and some mitigation plans have been implemented locally, the study of the interactions of P. miles with Mediterranean species and their impact on the local biodiversity is in its infancy. Closing this gap will lead to important fundamental insights in invasion ecology and will result in predictions on the impact of P. miles on the ecology and ecosystem services of the Mediterranean Sea. Such information will have practical implications for policy-makers aiming to devise sound and efficient mitigation plans.

Numerical Investigation of Some Reductions for the Gatenby–Gawlinski Model

Two (consecutive) reductions of the complete Gatenby–Gawlinski model for cancer invasion are proposed in order to investigate the mathematical framework, mainly from a computational perspective. After a brief overview of the full model, we proceed by examining the case of a two-equations-based and one-equation-based reduction, both obtained by means of a quasi-steady-state assumption. We focus on invasion fronts, exploiting a numerical strategy based on a finite volume approximation, and perform corresponding computational simulations to study the sharpness/smoothness of the traveling waves. Then, we employ a space-averaged wave speed estimate—referred to as the LeVeque–Yee formula—to quantitatively approach the propagation phenomenon. Concerning the one-equation-based model, we propose a scalar degenerate reaction-diffusion equation, which proves to be effective in order to qualitatively recover the typical trends arising from the Gatenby–Gawlinski model. Finally, we carry out some numerical tests in a specific case where the analytical solution is available.

Chordoma cells possess bone-dissolving activity at the bone invasion front.

Chordomas are malignant tumors that destroy bones, compress surrounding nerve tissues and exhibit phenotypes that recapitulate notochordal differentiation in the axial skeleton. Chordomas recur frequently, as they resist radio-chemotherapy and are difficult to completely resect, leading to repeated bone destruction and local expansion via unknown mechanisms. Here, using chordoma specimens and JHC7 chordoma cells, we asked whether chordoma cells possess bone-dissolving activity. CT imaging and histological analysis were performed to evaluate the structure and mineral density of chordoma-invaded bone and osteolytic marker expression. JHC7 cells were subjected to immunocytochemistry, imaging of cell fusion, calcium dynamics and acidic vacuoles, and bone lysis assays. In patients, we found that the skull base invaded by chordoma was highly porous, showed low mineral density and contained brachyury-positive chordoma cells and conventional osteoclasts both expressing the osteolytic markers tartrate-resistant acid phosphatase (TRAP) and collagenases. JHC7 cells expressed TRAP and cathepsin K, became multinucleated via cell-cell fusion, showed spontaneous calcium oscillation, and were partly responsive to the osteoclastogenic cytokine RANKL. JHC7 cells exhibited large acidic vacuoles, and nonregulatory bone degradation without forming actin rings. Finally, bone-derived factors, calcium ions, TGF-β1, and IGF-1 enhanced JHC7 cell proliferation. In chordoma, we propose that in addition to conventional bone resorption by osteoclasts, chordoma cells possess bone-dissolving activity at the tumor-bone boundary. Furthermore, bone destruction and tumor expansion may occur in a positive feedback loop.

Interleukin-6-Positive Immune Cells as a Possible New Immunologic Marker Associated With the Colorectal Cancer Prognosis.

Chronic inflammation creates tumor microenvironment (TME) that facilitates colorectal cancer (CRC) cell proliferation, migration, metastasis, and tumor progression. Interleukin-6 (IL-6) is a proinflammatory cytokine with a pleiotropic effect on CRC development. We aimed to evaluate IL-6 expression in tumor cells and in immune cells in TME, to assess the serum level and IL6 -174 G/C genotype distribution and to correlate the results with selected morphologic and clinical parameters that may add useful information in understanding the mechanisms of human CRC progression. A total of 153 patients with CRC were recruited in the current study. We assessed the IL-6 serum concentration through the ELISA method, the expression of IL-6 in tumor and in immune cells by immunohistochemical and double immunofluorescence staining, the MSI status by immunоhistochemistry for 4 mismatch repair (MMR) proteins, and the genotype distributions for IL6 -174G/C (rs1800795) single-nucleotide polymorphism through PCR-RFLP method. Our results showed that serum IL-6 level were increased in CRC patients as compared with healthy controls (P<0.0001), and in patients with cancers with advanced histologic type (type IV). However, the higher concentration (above the median of 55.71pg/mL) was with borderline association with longer survival of the patients after surgical therapy (P=0.055, Log rank test). We also found that IL-6+ immune cells prevailed in the invasive front (IF) of tumors compared with the tumor stroma (TS) (P<0.0001). More IL-6+ cells were recruited in the tumors with less advanced histologic type (I+II), with stronger inflammatory infiltrate in the IF, in early pTNM stages (I+II), without lymph node and distant metastases and the higher levels of IL-6+ cells, especially in the IF, were associated with longer survival (P=0.012). The results of our study suggest that although the serum levels of IL-6 are higher in CRC, the increased IL-6+ cells in tumor microenvironment, both in the invasive front and in tumor stroma, as well as the higher serum levels are associated with good prognostic variables and longer survival of the patients mainly in the early stages of CRC.

Drivers of Hymenoscyphus fraxineus Infections in the Inner-Alpine Valleys of Northwestern Italy

Fraxinus excelsior L. (ash) is a key forest tree species challenged by Hymenoscyphus fraxineus (T. Kowalski) Baral, Queloz, Hosoya, the causal agent of ash dieback. The goals of this study were (I) to assess the presence, spatial distribution, and incidence of H. fraxineus in the inner-alpine valleys of northwestern Italy, along with the severity of ash dieback; (II) to model the probability of infection by H. fraxineus based on environmental variables; (III) to reconstruct the direction of provenance of the front of invasion of the pathogen; and (IV) to test whether H. fraxineus has replaced the native relative Hymenoscyphus albidus (Gillet) W. Phillips, a saprobe of ash litter. By combining phytosanitary monitoring and samplings in 20 forest stands, laboratory analyses, and statistical modelling, this study showed that H. fraxineus was present in 65% of stands with an average incidence of 27%, reaching peaks of 80%. Rainfalls were the most relevant drivers of the probability of infection by H. fraxineus, rising up to 80% with the increased precipitation in April and July. Other drivers included elevation, maximal temperatures, latitude, and longitude. The front of invasion likely moved from Italy and/or Switzerland, rather than from France, while the replacement of H. albidus is uncertain.

Misapplied management makes matters worse: Spatially explicit control leverages biotic interactions to slow invasion.

A wide range of approaches has been used to manage the spread of invasive species, yet invaders continue to be a challenge to control. In some cases, management actions have no effect or may even inadvertently benefit the targeted invader. Here, we use the mid-20th century management of the Red Imported Fire Ant, Solenopsis invicta, in the US as a motivating case study to explore the conditions under which such wasted management effort may occur. Introduced in approximately 1940, the fire ant spread widely through the southeast USand became a problematic pest. Historically, fire ants were managed with broad-spectrum pesticides; unfortunately, these efforts were largely unsuccessful. One hypothesis suggests that, by also killing native ants, mass pesticide application reduced competitive burdens thereby enabling fire ants to invade more quickly than they would in the absence of management. We use a mechanistic competition model to demonstrate the landscape-level effects of such management. We explicitly model the extent and location of pesticide applications, showing that the same pesticide application can have a positive, neutral, or negative effect on the progress of an invasion, depending on where it is applied on the landscape with respect to the invasion front. When designing management, the target species is often considered alone; however, this work suggests that leveraging existing biotic interactions, specifically competition with native species, can increase the efficacy of management. Our model not only highlights the potential unintended consequences of ignoring biotic interactions, but also provides a framework for developing spatially explicit management strategies that take advantage of these biotic interactions to work smarter, not harder.

Combined spatial Keratin expression profiles at the invasive front represent a prognostic classifier for head and neck cancer

Keratin proteins are intermediate filaments that provide mechanical support and guide a plethora of important functions in epithelial cells. Although Keratins can be effectively used to characterize and differentiate cell lineages in many epithelia, it is still unclear if combinations of Keratin types and their spatial expression patterns can aid prognostication of head and neck squamous cell carcinoma (HNSCC). Here, we have assessed a panel of Keratin markers in four HPV negative HNSCC patient-derived organoid (PDO) models and their corresponding primary patient tissue to define spatial expression profiles of basal and supra-basal Keratins during invasion in Collagen-I matrices. Based on these profiles we have devised a scoring classification consisting of the following profiles: (i) no expression (Negative), (ii) uniform expression (Homogenous), (iii) heterogeneous expression (Mosaic), (iv) expression in the tumor core (Core), and (v) expression at the invasive front (Edge). Subsequently, this 5-tier system was validated in a well-documented tissue cohort of 157 HNSCC patients. We observe that patients with a Keratin 13 (K13) Edge profile have a significantly better prognosis than patients with K13 Core expression or K13 Negative tumors. Interestingly, we also find that K14 mosaicism strongly correlates with a favorable prognosis. We show that the absence of K13 from the tumor edge, in combination with K14 homogenous expression is a strong biomarker for poor prognosis in HNSCC. In short, our work indicates that defining the spatial expression patterns of basal and supra-basal markers in invasive HNSCC can benefit patient prognostication.

Multi-scale effects of behavioral movement deterrents on invasive carp metapopulations

Behavioral deterrents of among-pool movement represent a promising tool for controlling invasive fish populations. To date, much of the research in this area has been focused on the direct effectiveness of different methods of deterrence. However, the effect of these structures on populations in spatially complex habitats is unknown. We combine a metacommunity model with movement data of two invasive species (bighead carp and silver carp) in a large river to assess local and river-wide scale population outcomes of deterrent locations. We calculated (1) which potential deterrent locations are most effective at reducing the growth at the invasion front (2) the river-scale population effects at each location, and (3) what, if any, are the risks imposed by altering the current spatial dynamics. We found that the effects on the population dynamics at the invasion front varied with the location of deterrents, ranging from near zero to effects equal to the reduction in an individual’s movement across the deterrent. The river-scale population growth rate was slightly increased by all potential deterrent placements because the deterrents tended to concentrate more of the river-scale population into pools with the highest recruitment rates. The short-term, transient dynamics followed a strictly decreasing pattern after deterrent placement suggesting no additional short-term risk. These results suggest that deterrents can be an effective and low-risk intervention for the control of invasive carp, although the population level effect will depend on the interaction of the traits and behavior of the species with the physical character and spatial structure of the habitat.

A Cross-Sectional Study of Tumor Budding in Correlation with Clinicopathological Parameters in Invasive Breast Carcinoma

Introduction: Breast carcinoma is the largest cause of mortality from cancer among women in India. Tumor budding is usually defined as isolated single cancer cells or clusters of up to four cancer cells located at the invasive tumor front. Tumor budding is associated with poor clinicopathological characteristics, such as Tumor size, tumor differentiation, lymph node invasion, lymphatic or vascular invasion etc. Aim: To study tumor budding and its association with known clinicopathological parameters and hormone receptor status in invasive breast carcinoma. Methods : A Cross-sectional study was conducted after approval from the ethics committee. The duration of study was 1.5 years (Jan 2022- June 2023). Results: A total of fifty cases of invasive breast carcinomas were included in this study. 70% (35cases) were &gt;50yrs of age, 65% (33cases) were of T2 (tumor size) category. 82%(41cases) of Invasive duct carcinoma, No specific type, 12% (6 cases) of mixed carcinomas followed by three cases, one is invasive lobular carcinoma(6%), others are high grade papillary duct carcinoma(6%) and mucinous carcinoma (6%). 76%(38cases) showed presence of lymphovascular invasion, 74%(37cases) showed absence of necrosis, 54%(27 cases) are categorized as grade 2. High grade tumor budding was seen in 54% (27 cases). Conclusion: The following variables had a significant association with the tumor budding in present study: Tumor size, TNM staging, treatment status, lymphovascular invasion showing a p-value of &lt;0.05. ER positive tumors are also associated with high grade tumor budding. Keywords: Breast Carcinoma, Clinicopathological Variables, Lymphovascular Invasion, Tumor Budding

Teacher toads: Buffering apex predators from toxic invaders in a remote tropical landscape

AbstractEven after research identifies new approaches for wildlife management, translating those methods for delivery can be logistically challenging. In tropical Australia, invasive cane toads (Rhinella marina) fatally poison many anuran‐eating native predators. Small‐scale trials show that vulnerable predators exposed to small (nonlethal) toads can learn to delete toads from their diets, increasing survival of those predators after toads invade. We deployed this method in the remote Kimberley region of tropical Australia, with &gt;200,000 eggs, tadpoles, or metamorph toads released in advance of the expanding toad invasion front. Remote camera surveys before and after “teacher toad” deployment showed that large monitor lizards (Varanus panoptes) were almost extirpated from control plots but remained abundant in treatment plots, indicating broad‐scale success of this novel intervention.

294 Multi-omic Screening of Invasive GBM Cells in Engineered Biomaterials and Patient Biopsies Reveals Targetable Transsulfuration Pathway Alterations

INTRODUCTION: The poor prognosis of glioblastoma (GBM) is driven by tumor invasion; however, little is known of the metabolic alterations that drive invasion in glioblastoma. METHODS: We integrated spatially addressable hydrogel biomaterial platforms, patient site-directed biopsies, and multi-omics analyses to define metabolic drivers of invasive glioblastoma cells. A CRISPR metabolic gene screen was utilized to identify metabolic genes crucial for GBM invasion. We further targeted an identified hit from our high-throughput screens by utilizing a sgRNA to generate a knockdown cell line and assessing invasion in vitro and in vivo in a patient-derived xenograft orthotopic mouse model. RESULTS: Metabolomics and lipidomics revealed elevations in the redox buffers cystathionine, hexosylceramides, and glucosyl ceramides in the invasive front of both hydrogel-cultured tumors and patient site-directed biopsies, with immunofluorescence indicating elevated reactive oxygen species (ROS) markers in invasive cells. Transcriptomics confirmed upregulation of ROS-producing and response genes at the invasive front in both hydrogel models and patient tumors. Amongst oncologic ROS, hydrogen peroxide specifically promoted glioblastoma invasion in 3D hydrogel spheroid cultures. A CRISPR metabolic gene screen revealed cystathionine gamma lyase (CTH), an enzyme which converts cystathionine to the non-essential amino acid cysteine in the transsulfuration pathway, to be essential for glioblastoma invasion. Correspondingly, supplementing CTH knockdown cells with exogenous cysteine rescued invasion. Pharmacologic CTH inhibition suppressed glioblastoma invasion, while CTH knockdown slowed glioblastoma invasion in vivo. CONCLUSIONS: Our studies highlight the importance of ROS metabolism in invasive glioblastoma cells and support further exploration of the transsulfuration pathway as a mechanistic and therapeutic target.

Habitat alteration or climate: What drives the densities of an invading ungulate?

Anthropogenic habitat alteration and climate change are two well-known contributors to biodiversity loss through changes to species distribution and abundance; yet, disentangling the effects of these two factors is often hindered by their inherent confound across both space and time. We leveraged a contrast in habitat alteration associated with the jurisdictional boundary between two Canadian provinces to evaluate the relative effects of spatial variation in habitat alteration and climate on white-tailed deer (Odocoileus virginianus) densities. White-tailed deer are an invading ungulate across much of North America, whose expansion into Canada's boreal forest is implicated in the decline of boreal caribou (Rangifer tarandus caribou), a species listed as Threatened in Canada. We estimated white-tailed deer densities using 300 remote cameras across 12 replicated 50 km2 landscapes over 5 years. White-tailed deer densities were significantly lower in areas where winter severity was higher. For example, predicted deer densities declined from 1.83 to 0.35 deer/km2 when winter severity increased from the lowest value to the median value. There was a tendency for densities to increase with increasing habitat alteration; however, the magnitude of this effect was approximately half that of climate. Our findings suggest that climate is the primary driver of white-tailed deer populations; however, understanding the mechanisms underpinning this relationship requires further study of over-winter survival and fecundity. Long-term monitoring at the invasion front is needed to evaluate the drivers of abundance over time, particularly given the unpredictability of climate change and increasing prevalence of extreme weather events.

Analyzing Tumor Budding Scores in Invasive Breast Carcinoma: A Tertiary Care Center Study in South India

ABSTRACT Introduction and Aim: Tumor budding is a distinctive phenomenon which involves the presence of small clusters or individual cancer cells at the invasive front of tumors. Tumor budding has garnered attention due to its potential implications for prognosis, treatment strategies, and our understanding of cancer progression. Our aim is to study the distribution of tumor buds and its scoring in patients with infiltrating breast carcinoma and to associate with other histopathological parameters like the size of the tumor, its grade, lymphovascular invasion, and lymph node metastasis. Materials and Methods: This was a study analyzing the data of 70 resected specimens of primary breast carcinomas and providing a descriptive overview. Tumor budding was recognized, counted, and graded in hematoxylin and eosin slides. The cases were classified as low (0–4), intermediate (5–9), and high (≥10 buds) based on the count of tumor buds. Tumor budding has significant correlation with tumor grade and tumor size. Results: Of the 70 cases, 60 cases (85.71%) were diagnosed as invasive ductal carcinoma NOS. The majority [38 (54.28%)] of the cases showed an intermediate tumor budding score of 5–9/10 HPF. Conclusion: Evaluation of tumor budding allows pathologists and oncologists to gather valuable information about the tumor’s biological aggressiveness and potential for metastasis. It also helps in better risk stratification of patients, enabling a more personalized and tailored approach to treatment planning. In conclusion, assessing tumor budding in breast carcinoma holds significant clinical importance in the management and prognosis of this disease.

Abstract 6273: GGCT expression is a useful marker for the diagnosis of follicular thyroid carcinoma

Abstract Background: Follicular thyroid carcinoma (FTC) and follicular adenoma (FA) have similar histological findings. FTC is distinct from FA based on the presence of capsular or vascular invasion in the tissues, but it is difficult to confirm the diagnosis in some cases. In this study, we aimed to establish useful molecular maker for differential diagnosis between FTC and FA. Methods: This study included FTC (n = 32), FA (n = 64), and follicular tumors of uncertain malignant potential (FT-UMP, n = 5). Invasive front tissue and tumor center tissue from 3 cases of FTC were collected using a laser microdissection system; RNA was extracted, followed by microarray analysis to examine molecules that were commonly highly expressed in the invasive front. Subsequently, immunohistochemical staining of gamma-glutamyl cyclotransferase (GGCT) was performed. For assessment of GGCT expression, the intensity of the nuclear staining was stratified using a four-tiered scale (0 to 3) and percentage of positive cells (0-100%) in the hotspot. Multiplying intensity score and percent of positive cells, an expression score ranging from 0 to 300 was calculated. The Ki-67 labeling index was examined in 20 cases of FTC, 25 cases of FA, and 4 cases of FT-UMP from the same cohort. We counted at least 2,000 tumor cells in the hot spot for the assessment of Ki-67 labeling index. Results: Microarray analysis revealed GGCT exhibited high expression levels at the invasive front of FTC. In immunohistochemical assessment, GGCT expression scores were significantly higher in FTC than in FA (118.5 ± 51.4 vs. 57.3 ± 34.7, p &amp;lt; 0.0001). The median Ki-67 labeling index were 3.9% (range 0.2 - 16.1) for FTC and 2.4% (range 0.3 - 7.0) for FA. FTC showed a higher Ki-67 index than FA (Wilcoxon rank sum test, p = 0.0444). With GGCT expression score, applying a cutoff value of 101.1, the distinction between FTC and FA resulted in a sensitivity of 68.8% and a specificity of 87.5% (AUC: 0.832). With Ki-67 labeling index, applying a cutoff value of 4.0%, the distinction between FTC and FA resulted in a sensitivity of 50.0% and a specificity of 80.0% (AUC: 0.677). The GGCT expression score was positively related with the Ki-67 labeling index in the FTC cases. (Spearman's ρ = 0.5293, p = 0.0164). There were no significant associations between GGCT expression scores and clinicopathological parameters, including age, gender, tumor diameter, pT stage, pN stage, pattern of capsular invasion, number of foci of vascular invasion, and the presence of distant metastasis. Conclusions: GGCT is a potential marker for differentiating FTC from FA. The GGCT expression of FTC may contribute to estimating the invasive activity and proliferative activity. Citation Format: Toshiyuki Mitsuhashi, Sachiko Ogasawara, Masamichi Nakayama, Reiichiro Kondo, Jun Akiba, Kenta Murotani, Takeharu Ono, Fumihiko Sato, Hirohito Umeno, Hirohisa Yano. GGCT expression is a useful marker for the diagnosis of follicular thyroid carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6273.

Abstract 6801: Cellular senescence involves spatial evolution in colorectal cancer

Abstract Cancer evolution is a continuous process involving cellular changes that promote progression and metastasis. Recent studies demonstrate that cancer evolution is spatially associated: a phenomenon called “the spatial evolution of cancer.” In our data, colorectal cancer cells gradually evolve from an adenoid to a collective invasion morphology, ultimately developing a partial epithelial-mesenchymal transition phenotype as they form an invasive front. Trajectory analysis showed that cancer cells at the invasive front of a tumor exhibit a totally different gene expression pattern from those at the center, and that cellular senescence plays a crucial role in this transition. Epigenetic changes in the promoter/exon1 region of the CDKN2A gene caused by increased reactive oxygen species level are crucial for expression of p16INK4A and induction of senescence in the cancer cells. Two types of senescent tumor cells (type I and type II) were observed, and they played different roles in cancer progression due to different gene expression. Type II senescent tumor cells (p16INK4A+/LAMC2+/MMP7+), a final evolved form of cancer cells, are strongly associated with local invasion and lymph node metastasis of colorectal cancer and worsen the prognosis for patients. Citation Format: Tae Jun Park, Seok Yun Kang, Jang Hee Kim, Hyun Woo Lee, Yong Won Choi, Soon Sang Park, Young-Kyoung Lee. Cellular senescence involves spatial evolution in colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6801.

Abstract 7666: Type XVII collagen as potential biomarker of epithelial damage in patients with atopic dermatitis and patients with metastatic colorectal cancer

Abstract Background Type XVII collagen is a membrane-associated collagen with interrupted triple-helices (MACIT) facilitating adhesion between the basement membrane (BS) and epithelial cells. Mutations in COL17A1 lead to junctional epidermolysis bullosa (JEB), while autoimmunity against type collagen XVII ectodomain can cause bullous pemphigoid (BP). In multiple epithelial cancers, both type XVII collagen and its shed ectodomain are associated with cancer development and invasiveness. This study aims to assess the potential of type XVII collagen as an epithelial damage biomarker in patients with metastatic colorectal cancer (mCRC) and in patients with skin diseases like atopic dermatitis (AD) and psoriasis. Methodology Type XVII collagen ectodomain levels were measured in serum using a competitive ELISA (nordicPRO-C17). The study included two cohorts: cohort 1 comprised 212 patients with mCRC and 28 healthy controls and cohort 2 included patients with AD (n=8) or psoriasis (n=3). PRO-C17 levels in mCRC patients were compared to 28 healthy controls using a t-test. The association between PRO-C17 levels and overall survival (OS) was evaluated by categorizing patients into low PRO-C17 levels (tertiles 1+2) and high PRO-C17 levels (tertile 3). Kaplan-Meier curves and the log-rank test were used to assess differences in survival between the two groups. The independent predictive value of PRO-C17 was evaluated using multivariate Cox analysis. PRO-C17 levels in AD and psoriasis patients were compared to 15 healthy controls using one-way ANOVA, followed by multiple comparisons with Dunn's test. Results PRO-C17 levels were elevated (p&amp;lt;0.05) in mCRC patients treated with bevacizumab compared to healthy controls. Patients with high PRO-C17 levels had a shorter overall survival (390 days) compared to those with low levels (539 days) (p=0.007). High levels of the biomarker were associated with an increased risk of mortality and PRO-C17 levels were independent of other risk factors, such as the number of drugs in the treatment regimen, tumor location, synchronous metastatic disease, primary tumor resection, performance status, and the number of metastatic sites (HR=1.43, 95% CI 1.01-2.01, p=0.044). Further, patients with AD had increased PRO-C17 levels (p=0.0057), while patients with psoriasis showed no significant differences in biomarker levels compared to healthy controls (p=0.58). Conclusions This study demonstrated that high PRO-C17 levels were independently associated with shorter OS and an increased risk of mortality in patients with mCRC. Additionally, elevated levels of PRO-C17 were observed in patients with AD. These findings suggest that type XVII collagen may serve as a reflection of epithelial damage in the invasive front during tumor metastasis. The role of this biomarker in other cancer types and skin diseases warrants further exploration. Citation Format: Marina Crespo-Bravo, Neel Nissen, Rasmus Pedersen, Mogens Boisen, Maria Liljefors, Astrid Johansen, Julia Johansen, Morten Karsdal, Nicholas Willumsen. Type XVII collagen as potential biomarker of epithelial damage in patients with atopic dermatitis and patients with metastatic colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7666.

Abstract 5156: Is the formation of Stroma AReactive Invasion Front Areas in colorectal cancer driven by characteristic extracellular matrix reorganization processes based on the plasmin/plasminogen system

Abstract Background: As patient stratification still needs to be refined in colorectal cancer (CRC), we established SARIFA (Stroma AReactive Invasion Front Areas) as a novel H&amp;E-based prognostic biomarker that is defined by a direct tumor-adipocyte interaction with a lack of desmoplastic reaction. Even though we could prove the high prognostic relevance of SARIFA now in multiple cohorts in gastric, colorectal, and pancreatic cancer, as well as could link an upregulation of lipid metabolism and immune alterations to this specific aggressive tumor biology, it remains an open question how SARIFAs are formed mechanistically. Materials/Methods: Based on the results of our recently established SARIFA-scores in the TCGA-CRC (TCGA-COAD &amp; TCGA-READ cohorts; SARIFA-positive n=69, SARIFA-negative n=138) and the corresponding differential gene expression analysis, which are part of a previous study, protein levels (in ng/mL) measured by commercially available uPA and PAI1 enzyme-linked immunosorbent assays (ELISAs) were compared between SARIFA-positive and SARIFA-negative CRC patients (SARIFA-positive n=35, SARIFA-negative n=62). Additionally, immunohistochemistry for FABP4 as a key player in lipid metabolism was performed. Moreover, spatial transcriptomics (digital spatial profiling, DSP) was performed with tumor and stroma section segmentation on a subset of cases. Results: In TCGA-CRC, SERPINE1 (PAI1), as well as PLAU (uPA), are both significantly upregulated on mRNA level (LFC: 1.14 and 0.74, respectively; both adjusted p-values ≤0.001). Logically, SERPINE1 and PLAU are highly expressed in mesenchymal CMS4 CRCs, which overlap with SARIFA-positivity. This is reflected by an upregulation of matrix metalloproteinases in predicted functional protein networks (STRING analysis). In our local cohort, we could now validate this upregulation of uPA in SARIFA-positive CRCs on protein level (p=0.0027). For PAI1, a trend towards higher protein levels in SARIFA-positive CRCs could be observed (p=0.209). Moreover, SARIFA-positive CRCs show a higher number of tumor buds (p=0.013) as well as increased FABP4 expression at the invasion front (p&amp;lt;0.001) in our local cohort. In DSP analysis, the stroma fraction of SARIFA-positive CRCs showed significant changes compared to SARIFA-negative cases. Conclusion: Our current data shows that the formation of SARIFAs and the consecutive direct tumor-adipocyte interaction with a tumor-promoting increase in lipid metabolism may rely on alterations in the plasmin/plasminogen system, which could be a novel potential drug target in the aggressive subset of SARIFA-positive CRCs. Citation Format: Bianca Grosser, Nic G. Reitsam, Florian Sommer, Jochen Hardt, Bruno Märkl. Is the formation of Stroma AReactive Invasion Front Areas in colorectal cancer driven by characteristic extracellular matrix reorganization processes based on the plasmin/plasminogen system [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5156.

Abstract 1155: Spatial colocalization and molecular crosstalk of myofibroblastic CAFs and tumor cells shape lymph node metastasis in oral squamous cell carcinoma

Abstract Lymph node metastasis (LNM) is a critical prognostic factor for patients with oral squamous cell carcinoma (OSCC). While previous research has implicated partial epithelial-to-mesenchymal transition (p-EMT) of tumor cells and cancer-associated fibroblasts (CAFs), specifically myofibroblastic CAFs (myCAFs), in LNM, the underlying molecular mechanisms remain poorly understood. Furthermore, the lack of diagnostic and prognostic biomarkers related to LNM has impeded the improvement in clinical outcomes for patients with OSCC. Here, we conducted a comprehensive molecular analysis integrating original and publicly available OSCC data of the bulk genome and transcriptome analyses, single-cell transcriptome profiling, and spatial transcriptome analyses. We found that myCAFs were both quantitatively and functionally activated in LNM-positive samples and spatially colocalized with OSCC cells within the invasive tumor front (ITF), providing a niche that may facilitate LNM. We validated spatial colocalization of myCAFs and OSCC cells using 50 additional Immunohistochemistry (IHC) samples. Mechanistically, myCAFs provided increased extracellular matrix (ECM) signals, such as COLLAGEN, Fibronectin1(FN1), and LAMININ to OSCC cells. Consequently, OSCC cells upregulate ECM receptors such as αβ integrins (ITGA-B), syndecans (SDC), and CD44, leading to upregulating stemness-related genes in OSCC cells. Further analysis using CRISPRGeneEffect data from DepMap found that ITGB1, ITGA3, and CD44 are among the most essential genes for the survival and growth of OSCC cells. By analyzing the spatial transcriptomic data of matched primary tumor sites (HUH001-P1 and HUH001-P2) and metastatic site (HUH001-met) from the same patient, we identified the origins of metastatic cells in the primary tumor site. We further extracted a 23-gene signature from the metastatic origin in ITF, where OSCC and myCAFs colocalize. This spatially-resolved 23-gene signature predicted LNM status and poor overall survival (OS) in clinical OSCC patients. The prognostic significance of the 23-gene signature we extracted from the spatial analysis was also validated in separate clinical cohorts from TCGA study and two other microarray datasets. Our findings provide novel insight into the molecular crosstalk between myCAFs and OSCC cells that facilitates LNM and identifies clinically valuable biomarkers for better prognostication in patients with OSCC. Citation Format: Ken Furudate, Shuya Kasai, Tadashi Yoshizawa, Yuya Sasaki, Kohei Fujikura, Shintaro Goto, Ryohei Ito, Tomoyuki Tanaka, Hiroshi Kijima, Kosei Kubota, Ken Itoh, Wataru Kobayashi, Koichi Takahashi. Spatial colocalization and molecular crosstalk of myofibroblastic CAFs and tumor cells shape lymph node metastasis in oral squamous cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1155.