Tumor Edge Research Articles

Prognostic Value of CD8+ Lymphocytes in Hepatocellular Carcinoma and Perineoplastic Parenchyma Assessed by Interface Density Profiles in Liver Resection Samples.

Hepatocellular carcinoma (HCC) often emerges in the setting of long-standing inflammatory liver disease. CD8 lymphocytes are involved in both the antitumoral response and hepatocyte damage in the remaining parenchyma. We investigated the dual role of CD8 lymphocytes by assessing density profiles at the interfaces of both HCC and perineoplastic liver parenchyma with surrounding stroma in whole-slide immunohistochemistry images of surgical resection samples. We applied a hexagonal grid-based digital image analysis method to sample the interface zones and compute the CD8 density profiles within them. The prognostic value of the indicators was explored in the context of clinicopathological, peripheral blood testing, and surgery data. Independent predictors of worse OS were a low standard deviation of CD8+ density along the tumor edge, high mean CD8+ density within the epithelial aspect of the perineoplastic liver-stroma interface, longer duration of surgery, a higher level of aspartate transaminase (AST), and a higher basophil count in the peripheral blood. A combined score, derived from these five independent predictors, enabled risk stratification of the patients into three prognostic categories with a 5-year OS probability of 76%, 40%, and 8%. Independent predictors of longer RFS were stage pT1, shorter duration of surgery, larger tumor size, wider tumor-free margin, and higher mean CD8+ density in the epithelial aspect of the tumor-stroma interface. We conclude that (1) our computational models reveal independent and opposite prognostic impacts of CD8+ cell densities at the interfaces of the malignant and non-malignant epithelium interfaces with the surrounding stroma; and (2) together with pathology, surgery, and laboratory data, comprehensive prognostic models can be constructed to predict patient outcomes after liver resection due to HCC.

Imaging of Indocyanine Green-Human Serum Albumin (ICG-HSA) Complex in Secreted Protein Acidic and Rich in Cysteine (SPARC)-Expressing Glioblastoma.

Glioblastoma is the most common and fatal primary glioma and has a severe prognosis. It is a challenge for neurosurgeons to remove brain tumor tissues completely by resection. Meanwhile, fluorescence-guided surgery (FGS) is a technique used in glioma surgery to enhance the visualization of tumor edges to clarify the extent of tumor resection. Indocyanine green (ICG) is the only FDA-approved NIR fluorescent agent. It non-covalently binds to human serum albumin (HSA). Secreted protein acidic and rich in cysteine (SPARC) is an extracellular glycoprotein expressed in gliomas and binds to albumin, suggesting that it plays an important role in tumor uptake of the ICG-HSA complex. Here we demonstrate the binding properties of HSA or SPARC to ICG using surface plasmon resonance and saturation binding assay. According to in vitro and in vivo studies, the results showed that the uptake of ICG-HSA complex was higher in SPARC-expressing glioblastoma cell line and tumor region compared with the uptake of free ICG. Here, we visualized the SPARC-dependent uptake of ICG and ICG-HSA complex in U87MG. Our results demonstrated that the ICG-HSA complex is likely to be used as an efficient imaging agent targeting SPARC-expressing tumors, especially glioblastoma.

Focal Boundary Dice: Improved Breast Tumor Segmentation from MRI Scan.

Focal Boundary Dice, a new segmentation evaluation measure, was hereby presented, with the focus on boundary quality and class imbalance. Extensive analysis was carried out across different error types with varied object sizes of imaged tumors from Magnetic Resonance Imaging (MRI) scans, and the results show that Focal Boundary Dice is significantly more adaptive than the standard Focal and Dice measures to boundary errors for imaged tumors from MRI scans and does not over-penalize errors on the division of the boundary, including smaller imaged objects. Based on Boundary Dice, the standard evaluation protocols for tumor segmentation tasks were updated by proposing the Focal Boundary Dice. The contradiction between the target and the background area, and the conflict between the importance and the attention of boundary features were mainly solved. Meanwhile, a boundary attention module was introduced to further extract the tumor edge features. The new quality measure presents several desirable characteristics, including higher accuracy in the selection of hard samples, prediction/ground-truth pairs, and balanced responsiveness with across scales, which jointly make it more suitable for segmentation evaluation than other classification-focused measures such as combined Intersection-over-Union and Boundary binary cross-entropy loss, Boundary binary cross-entropy loss and Shape-aware Loss. The experiments show that the new evaluation metrics allow boundary quality improvements and image segmentation accuracy that are generally overlooked by current Dice-based evaluation metrics and deep learning models. It is expected that the adoption of the new boundary-adaptive evaluation metrics will facilitate the rapid progress in segmentation methods, and further contribute to the improvement of classification accuracy.

Novel MRI deformation-heterogeneity radiomic features are associated with molecular subgroups and overall survival in pediatric medulloblastoma: Preliminary findings from a multi-institutional study.

Medulloblastoma (MB) is a malignant, heterogenous brain tumor. Advances in molecular profiling have led to identifying four molecular subgroups of MB (WNT, SHH, Group 3, Group 4), each with distinct clinical behaviors. We hypothesize that (1) aggressive MB tumors, growing heterogeneously, induce pronounced local structural deformations in the surrounding parenchyma, and (b) these local deformations as captured on Gadolinium (Gd)-enhanced-T1w MRI are independently associated with molecular subgroups, as well as overall survival in MB patients. In this work, a total of 88 MB studies from 2 institutions were analyzed. Following tumor delineation, Gd-T1w scan for every patient was registered to a normal age-specific T1w-MRI template via deformable registration. Following patient-atlas registration, local structural deformations in the brain parenchyma were obtained for every patient by computing statistics from deformation magnitudes obtained from every 5mm annular region, 0 < d < 60 mm, where d is the distance from the tumor infiltrating edge. Multi-class comparison via ANOVA yielded significant differences between deformation magnitudes obtained for Group 3, Group 4, and SHH molecular subgroups, observed up to 60-mm outside the tumor edge. Additionally, Kaplan-Meier survival analysis showed that the local deformation statistics, combined with the current clinical risk-stratification approaches (molecular subgroup information and Chang's classification), could identify significant differences between high-risk and low-risk survival groups, achieving better performance results than using any of these approaches individually. These preliminary findings suggest there exists significant association of our tumor-induced deformation descriptor with overall survival in MB, and that there could be an added value in using the proposed radiomic descriptor along with the current risk classification approaches, towards more reliable risk assessment in pediatric MB.

Prediction of World Health Organization /International Society of Urological Pathology (WHO/ISUP) Pathological Grading of Clear Cell Renal Cell Carcinoma by Dual-Layer Spectral CT

To evaluate whether the dual-layer spectral computed tomography urography (DL-CTU) images could predict WHO/ISUP pathological grading of clear cell renal cell carcinoma (ccRCC). We retrospectively included patients (n=50) with pathologically confirmed ccRCC who underwent preoperative DL-CTU (from October 2017 to February 2021). They were divided into low-grade (WHO/ISUP 1/2, n=30) and high-grade groups (WHO/ISUP 3/4, n=20). The lesion size, attenuation (HU), iodine concentration (IC), normalized IC(NIC), and other quantitative characteristics were compared between the two groups. HU, IC, and NIC were obtained by plotting ROI with two different methods (circular ROI in the solid component or irregular ROI along the tumor edge containing tumor necrotic components). Receiver operating characteristic curves and multivariable model were used to evaluate the ability of parameters to predict WHO/ISUP grade. Transverse diameter (TD) of low-grade tumors was smaller, and HU in the non-contrast phase of the second method (HU-U-2) was lower than that of high-grade tumors (34.21±15.14 mm vs. 46.50 ± 20.68 mm, 27.33 ± 6.65 HU vs. 31.36 ± 6.09 HU, p< 0.05). The NIC in the nephrographic phase by the two methods (NIC-N-1 and NIC-N-2) of low-grade was higher than that of the high-grade group (0.78± 0.19 vs.0.58 ± 0.22, 0.73 ± 0.42 vs. 0.46 ± 0.22, p< 0.05). The final multivariable model composed of TD, HU-U-2, and NIC-N-1 could predict ccRCC grade with the area under the curve, sensitivity, specificity, and accuracy of 0.852, 70%, 90%, and 82%. Quantitative indicators in DL-CTU images could help predict the WHO/ISUP grade of ccRCC.

Analysis of Prevention and Treatment of Anastomotic Leakage after Sphincter-Preserving Surgery for Middle- and Low-Grade Rectal Cancer under Laparoscopy.

Anastomotic leakage is one of the most serious complications that can occur after laparoscopic-assistedsphincter-preserving surgery for middle- and low-grade rectal cancer. To explore the cause, prevention, and treatment of anastomotic leakage after sphincter-preserving surgery for middle- and low-grade rectal cancer under laparoscopy. The clinical data from patients with mid- and low-grade rectal cancer who underwent laparoscopic-assistedanus-preserving surgery in the anorectal surgery department of our hospital have been analyzed. Patients with a definite diagnosis, indications for laparoscopic surgery, and sphincter-preserving surgery were included in the analysis, and patients with a protective loop ileostomy and laparotomy were excluded. Among the 126 patients with middle- and low-grade rectal cancer undergoing sphincter-preserving surgery under laparoscopy. There were 75 male patients and 51 female patients, ranging in age from 37 to 89 years old, with an average age of 60.2 ± 6.7. The distance from the lower edge of the rectal tumor to the anal edge was ≤10 cm. 6 developed anastomotic leakage after the operation (leakage rate of 4.7%). Moreover, turbid purulent fluid was drained from the abdominal drainage tube in three patients on the third and fourth days after the operation, and the abdominal drainage tube drained serous drainage in three more patients on the fifth and sixth days, with signs of peritonitis appearing locally. All patients received continuous flushing and negative pressure drainage with a self-made double cannula and symptomatic treatment, and all were cured and discharged. Many factors can cause anastomotic leakage after this operation, and adequate perioperative preparation, meticulous operation during surgery, and careful postoperative management are key factors in preventing it.

MMP-7 expression is associated with a higher rate of tumor spread through air spaces in resected lung adenocarcinomas

ObjectivesThe spread through air spaces (STAS) of adenocarcinoma (ADC) is a unique pattern for local invasion, which comprises the spread of tumor cells within air spaces beyond the tumor edge without a direct connection with the primary tumor. Matrix metalloproteinase-7 (MMP-7), a secreted proteolytic enzyme that degrades various extracellular matrix components and other substrates, regulates several pathophysiological processes as well as the occurrence and development of cancers in humans. Here, we retrospectively analyzed a cohort of Japanese patients with treatment-naive, surgically-resected lung ADC to assess whether MMP-7 is associated with STAS development and if it could be used as a predictor of STAS. Materials and methodsWe performed histological evaluation using hematoxylin and eosin staining and immunohistochemical analysis using microarrays. Thereafter, we scored the examined tissues for immune markers to identify significant tumor STAS predictors. ResultsWe identified that high MMP-7 expression is an independent predictor of a high STAS incidence. Multivariate analysis revealed that MMP-7 expression was correlated with tumor behavior and poor prognosis. Furthermore, STAS remained significantly associated with a higher risk of ADC recurrence. ConclusionThe development of tumor STAS could be promoted by the functioning of MMP-7. This study could be a crucial basis for future investigations on the detection of tumor STAS.

Potential of [11C](R)-PK11195 PET Imaging for Evaluating Tumor Inflammation: A Murine Mammary Tumor Model.

Breast tumor inflammation is an immunological process that occurs mainly by mediation of Tumor-Associated Macrophages (TAM). Aiming for a specific measurement of tumor inflammation, the current study evaluated the potential of Positron Emission Tomography (PET) imaging with [11C](R)-PK11195 to evaluate tumor inflammation in a mammary tumor animal model. Female Balb/C mice were inoculated with 4T1 cells. The PET imaging with [11C](R)-PK11195 and [18F]FDG was acquired 3 days, 1 week, and 2 weeks after cell inoculation. The [11C](R)-PK11195 tumor uptake increased from 3 days to 1 week, and decreased at 2 weeks after cell inoculation, as opposed to the [18F]FDG uptake, which showed a slight decrease in uptake at 1 week and increased uptake at 2 weeks. In the control group, no significant differences occurred in tracer uptake over time. Tumor uptake of both radiopharmaceuticals is more expressed in tumor edge regions, with greater intensity at 2 weeks, as demonstrated by [11C](R)-PK11195 autoradiography and immunofluorescence with TSPO antibodies and CD86 pro-inflammatory phenotype. The [11C](R)-PK11195 was able to identify heterogeneous tumor inflammation in a murine model of breast cancer and the uptake varied according to tumor size. Together with the glycolytic marker [18F]FDG, molecular imaging with [11C](R)-PK11195 may provide a better characterization of inflammatory responses in cancer.

Phrenic nerve injury after the percutaneous microwave ablation of lung tumors: A single-center analysis.

This study aimed to analyze the cases of phrenic nerve injury caused by the percutaneous microwave ablation of lung tumors conducted at our center and to explore the risk factors. The data of 455 patients who underwent the percutaneous microwave ablation of lung tumors at the Department of Interventional Radiology, First Affiliated Hospital of Fujian Medical University from July 2017 to October 2021, were retrospectively analyzed. The cases of phrenic nerve injury after the percutaneous ablation were reported to analyze the risk factors involved, such as the shortest distance between tumor margin and phrenic nerve, tumor size, and ablation energy. The groups were divided based on the shortest distance between the tumor edge and the phrenic nerve into group 1, d ≤ l cm; group 2, 1 < d ≤2 cm; and group 3, d >2 cm. Lesions with a distance ≤2 cm were compared in terms of tumor size and ablation energy. Among the 455 patients included in this study, 348 had primary lung cancer, and 107 had oligometastatic cancer. A total of 579 lesions were detected, with maximum diameter of 1.27 ± 0.55 cm, and the ablation energy was 9,000 (4,800-72,000) J. Six patients developed phrenic nerve injury, with an incidence of 1.32%. For these six patients, the shortest distance from the lesion edge to the phrenic nerve was 0.75 ± 0.48 cm, and the ablation energy was 10,500 (8,400-34,650) J. There were statistically significant differences in phrenic nerve injury among groups 1, 2, and 3 (P < 0.05). In patients with a distance (d) ≤ 2 cm, there were no significant differences in tumor diameter and energy between the phrenic nerve injury group and the non-injury group (P = 0.80; P = 0.41). In five out of six patients, the diaphragm level completely recovered to the pre-procedure state, and the recovery time of the phrenic nerve was 9.60 ± 5.60 months. Another one was re-examined 11 months after the procedure, and the level of the diaphragm on the affected side had partially recovered. Phrenic nerve injury is a rare but not negligible complication of thermal ablation and is more likely to occur in lesions with a distance ≤2 cm from the phrenic nerve.

Mapping single-cell transcriptomes in the intra-tumoral and associated territories of kidney cancer.

Tumor behavior is intricately dependent on the oncogenic properties of cancer cells and their multi-cellular interactions. To understand these dependencies within the wider microenvironment, we studied over 270,000 single-cell transcriptomes and 100 microdissected whole exomes from 12 patients with kidney tumors, prior to validation using spatial transcriptomics. Tissues were sampled from multiple regions of the tumor core, the tumor-normal interface, normal surrounding tissues, and peripheral blood. We find that the tissue-type location of CD8+ Tcell clonotypes largely defines their exhaustion state with intra-tumoral spatial heterogeneity that is not well explained by somatic heterogeneity. De novo mutation calling from single-cell RNA-sequencing data allows us to broadly infer the clonality of stromal cells and lineage-trace myeloid cell development. We report six conserved meta-programs that distinguish tumor cell function, and find an epithelial-mesenchymal transition meta-program highly enriched at the tumor-normal interface that co-localizes with IL1B-expressing macrophages, offering a potential therapeutic target.

ANGI-06. CRISPRI SCREEN USING 3D BIOMIMETICS TO IDENTIFY NOVEL MARKERS OF GLIOBLASTOMA INVASION

Abstract Despite the aggressive treatment regimen, patients with GBM have a very poor prognosis, with a median survival of less than 2 years. The main reason behind this poor survival is the infiltration of tumor cells into the surrounding normal tissue. These infiltrative cells escape both surgical resection and radiation therapy, leading to inevitable tumor recurrence. Current approaches have failed to recognize that infiltrating GBM cells extending beyond the tumor edge have evolved a unique adaptive cellular machinery due to local stressors in their microenvironment. Unfortunately, these cells at the invasive tumor front are often not the ones sampled in studies analyzing banked tumor tissue, Second, most studies of invasion have used two-dimensional (2D) culture systems coated with a thin layer of extracellular matrix proteins, which fail to capture the dimensionality, mechanics, and heterogeneity of GBM invasion. We addressed these limitations by using an advanced CRISPRi druggable genome screen to identify novel druggable markers of GBM invasion in a 3D engineered biomimetic model comprised of Hyaluronic Acid (HA) based matrix. To better understand druggable pathways underlying GBM invasion, we performed a large-scale CRISPRi screen with a sgRNA library that targets the druggable human genome (2,550 genes). In the primary screen, sgRNAs targeting 12 genes were enriched in the non-invasive fraction. Out of these 12 genes, 8 genes are associated with cellular metabolism or chromosome assembly. We then validated these 8 genes using small molecule inhibitors in our 3D-gel sphere cultures, from which we identified four druggable targets, SOD-1 and NDUFV2 associated with mitochondrial metabolism and AURKB and ACP1 related to centromere assembly and chromosome segregation. In follow-up to these novel findings linking mitochondrial metabolism and centromere assembly to GBM invasion, our next steps are to identify the mechanism linking these genes to GBM cell invasion followed by validation in vivo.

CTNI-01. EFFECT OF PRE-OPERATIVE STEREOTACTIC RADIOSURGERY ON BRAIN METASTASIS: ANALYSIS OF DNA AND RNA GENOMIC PROFILES FROM PHASE-II CLINICAL TRIAL NCT03398694

Abstract BACKGROUND With improved systemic therapy that has limited impact on the intracranial compartment, the incidence of brain metastasis (BM) from solid cancers is rising. Treatment varies based on presentation, however, for patients with &amp;lt; 4 symptomatic BMs current clinical practice involves surgical resection followed by stereotactic radiosurgery (SRS) to the resection cavity. Post-operative SRS is associated with increased risk of leptomeningeal disease (LMD) and local recurrence. We hypothesize that pre-operative SRS will decrease the incidence of LMD as well as local recurrence, and increase patient’s OS by delivering a lethal dose of radiation to tumor cells before they are disturbed by surgical resection. In a Phase II clinical trial (NCT03398694) we are treating patients with 1-4 symptomatic BMs with pre-operative SRS while collecting DNA and RNA sequencing data from core and peripheral edges of the resected tumor to examine the genomic effects of SRS on tumor. METHODS Post-SRS resected tumor specimens were divided into two groups: ‘center’ and ‘periphery’ with respect to the center of SRS treatment with periphery within 50% isodose line. Previously resected untreated BMs were used as control. RESULTS/CONCLUSIONS Our initial data analyses show that pre-treatment with SRS results in significant genomic changes at DNA and RNA levels throughout the tumor when compared to non-radiated tumors. In matched center and peripheral samples DNA damage levels, measured by high impact mutation load, were similar indicating no evidence of radiation dose fall off effect on DNA damage. However, differing DNA damage locations and RNA expression profiles between center and matched samples are present, which may indicate unequal DNA repair processes between the center and periphery of tumors. Furthermore, global DNA mutation and RNA expression profiles differed significantly between the metastasis of different primary cancers signaling the importance of the primary cancer location in determining viable therapeutic targets.

DDDR-35. TARGETING GBM INVASION BY INHIBITING KCNA1 WITH 4-AMINOPYRIDINE: AN FDA APPROVED DRUG THAT EASILY PASS THROUGH THE BBB

Abstract Diffuse invasion is a hall mark of glioblastoma (GB) and one of the primary causes of poor clinical outcomes in GBM patients. Tumor cells migrate deep into the normal brain tissues are frequently protected by the BBB, making them particularly difficult to treat. New therapeutic targets are needed. Our previous studies using spatially dissected and functionally validated matching pairs of invasive and tumor core GBM cells identified KCNA1 as a shared gene that is selectively over-expressed in the invasive GBM cells in 6 patient derived orthotopic xenograft (PDOX) mouse models of pediatric GBM (Huang YL et al, Adv Science 2021). A subsequent analysis of adult GBM RNAseq data from IVY Atlas revealed a significantly elevated expression of KCNA1 (4.9 fold) in the invasive edges of patient GBM tumors. It is also one of the 11 core molecules identified in GEO and TCGA databases through an integrated bioinformatic analysis (Yang J, Front Onc 2021). To determine the anti-invasive activities of targeting KCNA1, we treated three highly invasive adult GBM PDOX models with 4-aminopyridine (4-AP), an old lipid soluble drug that easily penetrate the BBB, at 5 mg/kg, i.p., 5 days/week for 8 weeks acing alone or in combination with fractionated radiation (at 2 Gy/day x days). As single agent, 4-AP significantly extended median animal survival times in 1/3 GBM models (69 to 77 days, P = 0.033). Combination with XRT did not significantly improve the animal survival times in the three models. Systematic analysis of GBM invasion in mouse brains of the 3 PDOX models before, during and after 4-AP treatment revealed remarkable inhibition of tumor invasion. Our data highlighted the role of KCNA1 in promoting GBM invasion and support the fine tuning of 4-AP dose, schedule, and length of treatment to serve as a novel component of anti-GBM invasion therapies.

STEM-08. MODELING AND UNDERSTANDING GLIOBLASTOMA EDGE CELLS

Abstract Glioblastoma is a devastating disease with an overall median survival of 8 months from diagnosis. The majority of patients die of a tumor relapse in close proximity of the resected primary tumor. Glioblastoma is intensely researched but most studies have been performed on tissues and cultures derived from the bulk tumor while it is the remaining edge cells that cause lethality. Investigations of edge cells are rare and few experimental models exist. Here we have established and analyzed a series of matched cell cultures derived from the tumor bulk and outer edge of six IDH wildtype glioblastoma patients with the purpose to understand glioblastoma edge cell biology. Tumor samples were resected guided by 5-ALA fluorescence using neuro-navigation and stringent procedures to not contaminate edge samples with bulk tumor cells. First bulk tumor samples were resected from 5-ALA fluorescent tissue. After removal of all fluorescent areas and careful irrigation of the cavity the edge sample was resected 1-2 cm outside of the fluorescent border in a non-eloquent area. Following dissociation the samples were used in sphere assays and for explantation. There was a significant difference in self-renewal across all patients between matched bulk and edge cultures, in line with results from sphere assays on acute cells, suggesting maintenance of glioblastoma cell properties in established cultures. Invasion analysis showed a reverse significant difference between matched bulk and edge cultures strengthening a general functional distinction between tumor bulk and edge cells across patients. To investigate the molecular basis of our findings we performed whole exome sequencing (WES) and combined single cell RNA- and ATAC-sequencing (10X Multiome). Analyses are ongoing but WES data does not support genetic causes for their differences while the 10X Multiome data indicate that epigenetic regulation may underlie the different properties of bulk and edge glioblastoma cells.

NIMG-78. DEVELOPMENT OF A PRECLINICAL GLIOBLASTOMA MURINE MODEL FOR THE ASSESSMENT OF RADIATION NECROSIS USING DIFFUSION KURTOSIS IMAGING

Abstract Glioblastoma (GB) patients often present with either radiation-induced necrosis (RN) or develop tumor recurrence (TR). Radiologically, differentiating between these two disease states is particularly difficult, especially utilizing traditional MRI techniques. Accurately distinguishing these separate states is important, as failure to do so can ultimately lead to unnecessary surgical intervention or early cessation of appropriate radiation therapy. Currently, distinction between these pathologies is made using best clinical judgement by the neuro-oncology team. Therefore, there is a need for developing novel non-invasive techniques that can reliably distinguish between radiation necrosis and tumor recurrence. Primary data was collected utilizing orthotopically transplanted GL261 mouse GB cells in a C57BL/6 mouse. Tumor induction was verified by MRI after two weeks. RN and TR were initiated using an aggressive radiation dose fractionation (12 Gy or 60 Gy). This was completed using a 4 mm radiation cone such that one portion of the tumor received 100% dose of 12 Gy or 60 Gy fraction which is sufficient to cause RN, whereas the tumor edge received only 50% of the dose, allowing for tumor recurrence. Our data demonstrated mice tumor recurrence and radiation necrosis on MRI imaging. Axial T2-weighted MRI and DKI sequence at 2 weeks following implantation demonstrated edema compatible with tumor recurrence. One week later, the T2-weighted MRI and DKI sequences demonstrated continued tumor progression. In a separate mouse with orthotopic glioblastoma implantation with high dose (60 Gy) radiation treatment, T2 imaging and DKI demonstrated areas of tumor and suspected central radiation necrosis. DKI imaging biomarkers was compared by liquid biopsy, H&amp;E staining, and Immunohistochemistry (IHC) analysis of sacrificed mice. Our H&amp;E staining results showed typical pathological features of GB and RN, and TR in each case. This result demonstrated that our proposed model for radiation induction was effective at achieving its goal.

Auxiliary Segmentation Method of Osteosarcoma MRI Image Based on Transformer and U-Net.

One of the most prevalent malignant bone tumors is osteosarcoma. The diagnosis and treatment cycle are long and the prognosis is poor. It takes a lot of time to manually identify osteosarcoma from osteosarcoma magnetic resonance imaging (MRI). Medical image processing technology has greatly alleviated the problems faced by medical diagnoses. However, MRI images of osteosarcoma are characterized by high noise and blurred edges. The complex features increase the difficulty of lesion area identification. Therefore, this study proposes an osteosarcoma MRI image segmentation method (OSTransnet) based on Transformer and U-net. This technique primarily addresses the issues of fuzzy tumor edge segmentation and overfitting brought on by data noise. First, we optimize the dataset by changing the precise spatial distribution of noise and the data-increment image rotation process. The tumor is then segmented based on the model of U-Net and Transformer with edge improvement. It compensates for the limitations of U-semantic Net by using channel-based transformers. Finally, we also add an edge enhancement module (BAB) and a combined loss function to improve the performance of edge segmentation. The method's accuracy and stability are demonstrated by the detection and training results based on more than 4,000 MRI images of osteosarcoma, which also demonstrate how well the method works as an adjunct to clinical diagnosis and treatment.

STEM-02. GENERATION AND CHARACTERISATION OF A LIVING BIOBANK OF POST-SURGICAL RESIDUAL GLIOBLASTOMA TO IDENTIFY NOVEL THERAPEUTIC TARGETS

Abstract Glioblastoma is the most common cancer arising within the brain and contributes to ~190,000 brain tumour related deaths/year globally. Current standard-of-care therapy consists of maximal safe surgical resection, radiotherapy with concomitant then adjuvant chemotherapy (Temozolomide) which triggers the cellular DNA damage response (DDR). However, survival for patients remains poor (median OS ~ 12-15 months). Lack of progress in the development and clinical translation of novel therapeutics is largely attributed to extensive inter- and intra-tumoral heterogeneity and resistant glioma stem cell (GSC) niches. Traditionally, primary ex vivo and in vivo models of glioblastoma have been derived from the resected tumour mass potentially omitting significant genetic, epigenetic and phenotypical characteristics exhibited by post-surgical residual tumour cells that are ultimately responsible for disease recurrence and morbidity. We therefore hypothesise that GSC’s isolated from the invasive edge of the tumour are inherently treatment resistant due to an increased “stemness” and upregulated DDR. Here we display novel methodology for the generation of multiple parallel primary ex vivo GSC cell models from anatomically discrete regions of specific partial lobectomy specimens. Importantly, these samples contain large portions of infiltrated adjacent brain to recapitulate features of resected (tumour core) and typically residual glioblastoma (distal invasive tumour edge). Initial RNA-seq analysis has revealed large divergent transcriptional landscapes of resected vs residual tumour cells and protein expression analyses has identified key differences in DDR proteins and subsequent cellular responses to current standard-of-care radio-chemotherapy regimens. Immunofluorescence analysis of DNA damage markers has also revealed differential capacity &amp; kinetics for repair of DNA damage between resected and residual GSC populations. These data therefore highlight the potential clinical relevance of our novel parallel resected and residual ex vivo glioblastoma models, which will hopefully aid evaluation of novel therapeutics capable of targeting post-surgical GSC subpopulations leading to improved disease outcomes.

Liver tumor segmentation and classification using FLAS-UNet++ and an improved DenseNet.

The incidence of liver tumors is among the top three in China. The treatments of benign and malignant tumors are different. Accurate diagnosis plays an important role in guiding the treatment of tumors. The aim of this study is to solve the following: (1) blurred boundary between the liver tumor and other organs causes incorrect segmentation of liver tumor boundaries; (2) large difference in tumor size and the diversity in texture and grayscale are major challenges in liver tumor classification tasks. Firstly, the liver tumor is segmented from the original CT images by a tumor segmentation network, UNet++ with fusion loss and atrous spatial pyramid pooling (FLAS-UNet++). The proposed segmentation method can solve the problem of tumor edge segmentation error by learning the tumor edge information. Secondly they are adaptively cropped according to the tumor volume to reduce the over-fitting and over-sensitivity of the deep network. Thirdly an improved Dense Block is designed to pay more attention to the changes in grayscale and texture between benign and malignant tumors. Finally, the features extracted from the network combined with tumor volume, patient's sex and age, are sent to a classifier for diagnosis. Liver tumor segmentation results show that the dice, HD95 reached 71.9%, 12.1 mm, respectively. The classification results show that the accuracy, specificity, sensitivity and area under curve reached 82.4%, 79.8%, 84.4%, 87.5%, respectively. The segmentation and classification results are both better than other's methods and mainstream networks. In order to solve existing problems of liver tumor CT image classification methods, our method realizes the accurate segmentation and classification of liver tumors in CT images and has important clinical application value.

Mechanics of lung cancer: A finite element model shows strain amplification during early tumorigenesis.

Early lung cancer lesions develop within a unique microenvironment that undergoes constant cyclic stretch from respiration. While tumor stiffening is an established driver of tumor progression, the contribution of stress and strain to lung cancer is unknown. We developed tissue scale finite element models of lung tissue to test how early lesions alter respiration-induced strain. We found that an early tumor, represented as alveolar filling, amplified the strain experienced in the adjacent alveolar walls. Tumor stiffening further increased the amplitude of the strain in the adjacent alveolar walls and extended the strain amplification deeper into the normal lung. In contrast, the strain experienced in the tumor proper was less than the applied strain, although regions of amplification appeared at the tumor edge. Measurements of the alveolar wall thickness in clinical and mouse model samples of lung adenocarcinoma (LUAD) showed wall thickening adjacent to the tumors, consistent with cellular response to strain. Modeling alveolar wall thickening by encircling the tumor with thickened walls moved the strain amplification radially outward, to the next adjacent alveolus. Simulating iterative thickening in response to amplified strain produced tracks of thickened walls. We observed such tracks in early-stage clinical samples. The tracks were populated with invading tumor cells, suggesting that strain amplification in very early lung lesions could guide pro-invasive remodeling of the tumor microenvironment. The simulation results and tumor measurements suggest that cells at the edge of a lung tumor and in surrounding alveolar walls experience increased strain during respiration that could promote tumor progression.

Longitudinally Heterogeneous Tumor Dose Optimizes Proton Broadbeam, Interlaced Minibeam, and FLASH Therapy.

Simple SummaryThe aim of any kind of external radiation therapy is to control a tumor with the highest possible probability of the lowest possible side effects. Here, we study further opportunities of reducing the side effects of proton therapy by applying longitudinally heterogeneous dose distributions in the tumor respecting the delivery of a minimum prescribed dose. In our simulations, the longitudinally heterogeneous dose distributions show a reduced dose in the healthy tissue already in the case of proton broadbeam irradiations, but a much higher (calculated) mean cell survival in the case of proton minibeam irradiation. This demonstrates its potential to substantially reduce side effects at a simultaneously higher tumor control probability, opening new opportunities of easier application when striving for high dose-rate applications of proton beams (>~10 Gy/s), in order to additionally profit from the so-called FLASH effects.The prerequisite of any radiation therapy modality (X-ray, electron, proton, and heavy ion) is meant to meet at least a minimum prescribed dose at any location in the tumor for the best tumor control. In addition, there is also an upper dose limit within the tumor according to the International Commission on Radiation Units (ICRU) recommendations in order to spare healthy tissue as well as possible. However, healthy tissue may profit from the lower side effects when waving this upper dose limit and allowing a larger heterogeneous dose deposition in the tumor, but maintaining the prescribed minimum dose level, particularly in proton minibeam therapy. Methods: Three different longitudinally heterogeneous proton irradiation modes and a standard spread-out Bragg peak (SOBP) irradiation mode are simulated for their depth-dose curves under the constraint of maintaining a minimum prescribed dose anywhere in the tumor region. Symmetric dose distributions of two opposing directions are overlaid in a 25 cm-thick water phantom containing a 5 cm-thick tumor region. Interlaced planar minibeam dose distributions are compared to those of a broadbeam using the same longitudinal dose profiles. Results and Conclusion: All longitudinally heterogeneous proton irradiation modes show a dose reduction in the healthy tissue compared to the common SOBP mode in the case of broad proton beams. The proton minibeam cases show eventually a much larger mean cell survival and thus a further reduced equivalent uniform dose (EUD) in the healthy tissue than any broadbeam case. In fact, the irradiation mode using only one proton energy from each side shows better sparing capabilities in the healthy tissue than the common spread-out Bragg peak irradiation mode with the option of a better dose fall-off at the tumor edges and an easier technical realization, particularly in view of proton minibeam irradiation at ultra-high dose rates larger than ~10 Gy/s (so-called FLASH irradiation modes).