Metastatic Research Articles

HPB O02 - On-site hepatobiliary surgery capability independently predicts treatment pathway completion in metastatic rectal cancer: A nationwide observational study of 15,727 patients

Abstract Background Stage IV rectal cancer carries a 10% 5-year survival rate (Jemal et al., 2005). Curative-intent management requires treatment of both the primary and metastatic disease. This study aimed to identify patterns in stage IV rectal cancer treatment pathway at population level across the UK, and to identify predictors of treatment pathway completion. Method Patient demographics and treatment data were extracted from the UK Colorectal Cancer Intelligence Hub’s CORECT-R repository, national chemotherapy (SACT), radiotherapy datasets (RTDS), and Hospital Episode Statistics (HES) data. Treatment pathway completion was defined as receipt of intervention to the primary tumour, in addition to metastasis-directed therapy, or the combination alongside chemotherapy. Multivariable logistic regression analysis was performed to identify independent predictors of treatment completion. Results A total of 15,727 patients with stage IV rectal cancer diagnosed between 2010-19 were identified. There was significant variation in proportion of patients with complete treatment between MDTs. Multivariable analyses showed that lack of on-site HPB surgery capability was the only predictor of failure to complete treatment (OR 1.22, 95%CI:1.07-1.38, p<0.01). Conclusion Across the UK, there is variation in Stage IV rectal cancer patients completing treatment for both their primary and metastatic disease. Further work is needed to determine whether differences in treatment pathways stem from variation in access to specialist services between cancer alliances or whether they are due to differences in MDT decision making.

A Review of Limbic System-Associated Membrane Protein in Tumorigenesis

Purpose of Review: This review aims to describe the role of limbic system-associated membrane protein (LSAMP) in normal- and pathophysiology, and its potential implications in oncogenesis. We have summarized research articles reporting the role of LSAMP in the development of a variety of malignancies, such as clear cell renal cell carcinoma, prostatic adenocarcinoma, lung adenocarcinoma, osteosarcoma, neuroblastoma, acute myeloid leukemia, and epithelial ovarian cancer. We also examine the current understanding of how defects in LSAMP gene function may contribute to oncogenesis. Finally, this review discusses the implications of future LSAMP research and clinical applications. Recent Findings: LSAMP has been originally described as a surface adhesion glycoprotein expressed on cortical and subcortical neuronal somas and dendrites during the development of the limbic system. It is categorized as part of the IgLON immunoglobulin superfamily of cell-adhesion molecules and is involved in regulating neurite outgrowth and neural synapse generation. LSAMP is both aberrantly expressed and implicated in the development of neuropsychiatric disorders due to its role in the formation of specific neuronal connections within the brain. Additionally, LSAMP has been shown to support brain plasticity via the formation of neuronal synapses and is involved in modulating the hypothalamus in anxiogenic environments. In murine studies, the loss of LSAMP expression was associated with decreased sensitivity to amphetamine, increased sensitivity to benzodiazepines, increased hyperactivity in new environments, abnormal social behavior, decreased aggressive behavior, and decreased anxiety. Findings have suggested that LSAMP plays a role in attuning serotonergic activity as well as GABA activity. Given its importance to limbic system development, LSAMP has also been studied in the context of suicide. In malignancies, LSAMP may play a significant role as a putative tumor suppressor, the loss of which leads to more aggressive phenotypes and mortality from metastatic disease. Loss of the LSAMP gene facilitates epithelial-mesenchymal transition, or EMT, where epithelial cells lose adhesion and gain the motile properties associated with mesenchymal cells. Additionally, LSAMP and the function of the RTK pathway have been implicated in tumorigenesis through the modulation of RTK expression in cell membranes and the activation of second messenger pathways and β-catenin. Summary: Beyond its many roles in the limbic system, LSAMP functions as a putative tumor suppressor protein. Loss of the LSAMP gene is thought to facilitate epithelial-mesenchymal transition, or EMT, where cells lose adhesion and migrate to distant organs. LSAMP’s role in modulating RTK activity and downstream ERK and Akt pathways adds to a large body of data investigating RTK expression in oncogenesis. The characteristics of LSAMP defects and their association with aggressive and metastatic disease are evident in reports on clear cell renal cell carcinoma, prostatic adenocarcinoma, lung adenocarcinoma, osteosarcoma, neuroblastoma, acute myeloid leukemia, and epithelial ovarian cancer.

Abstract A038: Investigating PD-L1 status in circulating tumor cells isolated from blood samples of lung cancer patients

Abstract Background: This research study aimed to assess the performance of a research use only (RUO) immunofluorescence (IF) assay targeting programmed death-ligand 1 (PD-L1) evolution in epithelial and/or mesenchymal circulating tumor cells (CTCs) isolated from blood using Parsortix® microfluidic technology. Upregulation of PD-L1 enables cancer cells to evade the host immune response. Assessment of PD-L1 status in the tumor, as determined by traditional tissue biopsy, can indicate if immunotherapy has the potential to be an effective treatment. However, PD-L1 expression in tumor biopsies may not reflect metastatic sites in their entirety and can become outdated during tumor evolution, as the process cannot be repeated due its invasive nature. Liquid biopsy offers a minimally invasive option for dynamic testing of PD-L1 in CTCs as patients undergo treatment. Methods: Analytical linearity, and specificity and sensitivity were assessed by spiking HCC1954 and Hs 578T cancer cell lines into the blood samples of 34 healthy volunteers. The assay was then evaluated in samples from 47 metastatic lung cancer patients, among which 32 had a known PD-L1 tissue status (19 positive, 13 negative). Up to six successive draws were taken for each patient, collected into Streck Cell-Free DNA blood collection tubes. ANGLE’s Parsortix® instruments were used to isolate spiked cancer cells or CTCs from blood samples based on their size and deformability. The CTC- enriched harvests were processed onto ANGLE’s CellKeepTM slides and IF stained with ANGLE’s Portrait® PD-L1 RUO assay for PD-L1 identification on CTCs. Slides were imaged on a BioView automated imaging system. Results: Analytical specificity of PD-L1 was 98% and analytical sensitivity was 81%. Analytical linearity (R2=0.91) was shown over a 0–500 cells range. In metastatic lung cancer patients, ≥1 CTC was identified in at least one draw for 91% of donors, with 55% of those donors exhibiting ≥1 PD-L1 positive CTC. A majority (81%) of donors exhibited a positive correlation between PD-L1 tissue status and CTC expression. However, the remaining 19% of donors exhibited discordance with a PD-L1 negative tissue status but PD-L1 positivity identified in ≥1 CTC. Of the PD-L1 positive CTCs identified, 98% expressed mesenchymal markers. Dynamic change was shown in all but one donor, with PD-L1 status of the CTCs changing over time. Conclusions: This study demonstrated the ability to determine PD-L1 status in CTCs from the blood of metastatic lung cancer patients and, subject to further study, the future potential to develop dynamic PD-L1 testing for advancement of more personalized cancer treatments. Patients with a PD-L1 negative tissue status exhibited PD-L1 positive CTCs, demonstrating the feasibility of a more repeatable and accurate assessment of PD-L1 status than can be achieved through a traditional tissue biopsy. Citation Format: Morgan Spode, Chloe Goodwin, Aarabhi Varatharajah, Aaron Cottingham, Elisha Duhig, Laura Kaja, David Greaves, Mariacristina Ciccioli, Anne-Sophie Pailhes-Jimenez. Investigating PD-L1 status in circulating tumor cells isolated from blood samples of lung cancer patients [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr A038.

Associations of radiotherapy receipt with lung cancer risk by histological subtype among breast cancer survivors in the United States.

Radiotherapy for breast cancer has been associated with an increased risk of secondary malignancies, including primary lung cancer. Whether this association varies by histological subtype of lung cancer remains unknown. Based on the data from 12 Surveillance, Epidemiology, and End Results registries, we examined the association between radiotherapy receipt and the risk of subtype-specific subsequent primary lung cancer (SPLC) among female first primary breast cancer cases diagnosed between ages 20 and 84 from 1992 to 2020. More than half (53%) of the 550,007 breast cancer survivors identified had undergone radiotherapy as part of their initial breast cancer treatment. Over an average follow-up of 9.7 years, 8014 survivors developed SPLCs. For small-cell carcinoma, the standardized incidence ratio (SIR) compared with the general population was higher for survivors who received radiotherapy (SIR = 1.15, 95% confidence interval [CI] = 1.06-1.25) but similar for those who did not receive radiotherapy (SIR = 1.00, 95% CI = 0.91-1.09), with the difference in SIRs being statistically significant (p = .003). Similar associations were found for squamous cell carcinoma (SIRyes = 1.16, 95% CI = 1.08-1.24 vs. SIRno/unknown = 1.06, 95% CI = 0.98-1.15; p = .07). The increased risks were confined to ipsilateral SPLC, with the greatest SIRs for small-cell carcinoma occurring 5-10 years since breast cancer diagnosis (SIR = 1.83, 95% CI = 1.53-2.19) and for squamous cell carcinoma with a latency of 10 years or more (SIR = 1.64, 95% CI = 1.42-1.88). In contrast, the risk of developing adenocarcinoma did not vary by radiotherapy receipt (SIRyes = 1.23, 95% CI = 1.18-1.28 vs. SIRno/unknown = 1.17, 95% CI = 1.12-1.22; p = .18), indicating additional risk factors in play. The findings suggest a distinct carcinogenic pathway of radiation-induced lung cancer across histological subtypes and may inform risk-stratified surveillance guidelines for SPLC.

Abstract B042: Molecular assessment of cell surface targets using integrated circulating tumor cell (CTC) RNAseq and single CTC phenotyping

Abstract Background: Androgen-targeted therapies are a mainstay of treatment for metastatic prostate cancer and have significantly improved patient outcomes. However, development of treatment resistance remains universal, occurring through androgen receptor (AR) alterations driving constitutive AR signaling, or lineage state transitions that bypass AR entirely and culminate in a small cell/neuroendocrine phenotype (NEPC). Cell surface targeted therapies, including antibody drug conjugates and targeted radioligand therapies, represent a new therapeutic class that has shown promise for treatment of androgen-resistant metastatic prostate cancer (mCRPC). Circulating tumor cells (CTCs) present an accessible source of tumor material to identify expression of known and novel targets for therapeutic development, as well as the unique potential for longitudinal profiling to understand the evolution of target expression and association with clinical resistance to cell surface targeted therapies. We have developed a CTC platform allowing for gene expression and protein profiling of cell surface targets on CTCs. Methods: Live CTCs were isolated with our automated microfluidic technology integrating negative and positive selection for CTC enrichment with a cohort of 273 samples. CTCs were captured immunomagnetically followed by RNA isolation on chip and RNA-seq, or protein staining on chip for single cell fluorescent quantification of prostate adenocarcinoma and NEPC cell surface targets including PSMA, TROP2, B7H3 and DLL3. Results: CTC RNA sequencing can be used to understand the expression of cell surface targets across different CTC phenotypes. In CTCs from patient with mCRPC, canonical prostate adenocarcinoma cell surface targets STEAP1, KLK2 and FOLH1 and epithelial cell surface target TACSTD2 had the highest expression, while prostate adenocarcinoma target STEAP2, pan-tumor target ERBB2, and tumor immune checkpoint cell surface protein CD276 (B7H3) had intermediate expression. Expression of DLL3 and SSTR2, targets associated with neuroendocrine differentiation, was lowest, though still detected in a subset of CRPC CTCs. Comparison of expression levels between CRPC and NEPC CTCs demonstrated lower expression of most adenocarcinoma cell surface targets in NEPC CTCs as expected, and a trend towards higher expression of DLL3 and SSTR2. Cell surface expression is being assessed for proteins including Trop-2, PSMA and DLL3. Evaluation of DLL3 at the protein level in a subset of patients including both CRPC (n=13) and NEPC (n=12) confirmed that DLL3 was detected in both adenocarcinoma and neuroendocrine CTCs, with a median of 45.8% DLL3-positive CTCs per sample. Conclusions: Biomarkers are needed to better predict response and resistance to targeted therapies. CTC RNAseq and single CTC phenotyping hold potential for predictive and on-treatment biomarkers of response to cell surface targeted therapies. Further study of mechanisms driving response and resistance can be evaluated with these biomarkers. Citation Format: Jamie M. Sperger, Marina N. Sharifi, Katherine R. Kaufmann, Matthew L. Bootsma, Shannon R. Reese, Viridiana Carreno, Alex H. Chang, Luke A. Nunamaker A. Nunamaker, Charlotte Linebarger, Amy K. Taylor, Katherine E Tippins, Kyle T. Helzer, Shuang G. Zhao, Joshua M Lang. Molecular assessment of cell surface targets using integrated circulating tumor cell (CTC) RNAseq and single CTC phenotyping [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr B042.

Efficacy and safety of CalliSpheres drug-eluting bead bronchial arterial infusion chemoembolization vs. bland embolization in advanced lung cancer with hemoptysis: A multicenter retrospective study.

Massive hemoptysis is a life-threatening complication in patients with advanced primary lung cancer, and effective, safe treatments are crucial. This study aimed to investigate the efficacy and safety of CalliSpheres drug-eluting bead bronchial arterial infusion chemoembolization (DEB-BACE) for managing this condition. A retrospective analysis included 144 patients with advanced primary lung cancer and massive hemoptysis treated at multiple hospitals from January 2019 to January 2023. Patients undergoing bronchial artery embolization were divided into two groups: the observation group (n=76) received CalliSpheres DEB-BACE with epirubicin, and the control group (n=68) received 8spheres blank embolization. Both groups achieved successful hemostasis, with no statistically significant difference in success rates (observation group: 88.16%, control group: 86.76%). However, the observation group had a significantly longer median duration without hemoptysis (96 days vs. 50 days). Two months post-therapy, the observation group showed higher objective response rates (82.89% vs. 38.24%) and disease control rates (92.11% vs. 66.18%) compared to the control group. Adverse reactions were manageable and similar between groups, with no serious complications observed. By January 31, 2024, the observation group had significantly longer median overall survival (11 months vs. 7 months). The DEB-BACE treatment demonstrates safety and efficacy in managing massive hemoptysis in patients with advanced lung cancer. However, the superiority of this approach over bland embolization remains to be established through well-designed prospective studies. Future research is anticipated to provide a definitive comparison and further validate the role of DEB-BACE in clinical practice.

Abstract A026: A novel ddPCR detects hypermethylated RASSF1A in plasma of patients with pediatric Wilms tumor

Abstract Background: Wilms tumor is the most common pediatric renal cancer. Within the SIOP-Renal Tumor Study Group (SIOP-RTSG), diagnosis is based on imaging and patients receive neo- adjuvant treatment. Final diagnosis is established via histopathological analysis at the timepoint of surgery, which also determines the intensity of adjuvant treatment. Overall, patients with Wilms tumor have a 90% survival rate, but one in five patients will suffer from a relapse, which requires intensive treatment and induces long-term toxicity. It is challenging to identify patients at risk of suffering from relapse at initial diagnosis. Hence the need for additional biomarkers that will optimize risk stratification and minimal residual disease monitoring. RASSF1A is a tumor suppressor gene that is silenced via hypermethylation of its promotor region in multiple adult and pediatric tumors and is reported to represent an aggressive phenotype. We have previously reported on thirteen patients with Wilms tumor where increased hypermethylation of RASSF1A (RASSF1A-M) was observed in plasma cell-free DNA (cfDNA). The aim of this study was to further investigate the levels of RASSF1A-M in a larger cohort. Methods: The presence of RASSF1A-M in plasma cfDNA of patients with Wilms tumor was quantified using a multiplexed droplet digital PCR comprising methylation-specific restriction enzymes. Diagnosis and pre- surgery samples were included to evaluate RASSF1A-M levels during treatment. Results: A total of 49 patients were evaluated. At diagnosis, 37/46 (80.4%) samples were positive for RASSF1A-M. Negative samples were from patients with localized disease (N=6), regressive type regionally advanced Wilms (N=1) or mixed type metastatic Wilms (N=2). RASSF1A-M levels were significantly elevated in regionally advanced and metastatic disease compared to localized disease. Of the 37 positive diagnosis samples, 23 pre-surgical follow-up samples were available. In 7/23, RASSF1A-M was still detectable. These belong to patients with stromal-, regressive- or diffuse anaplastic predominant tumors. Interestingly, an increasing methylation level was seen in one patient with a regressive tumor. Conclusion: RASSF1A-M is a potential biomarker for patients with Wilms tumor, since it was detected in the majority of patients. Furthermore, significantly increased levels were observed in regionally advanced and metastatic disease. In the small number of patients with persisting levels of RASSF1A-M, we did not observe an association with risk groups. In collaboration with SIOP-RTSG, additional follow-up samples will be analyzed to explore the potential of RASSF1A-M as biomarker for relapse detection. Citation Format: Julia Sprokkerieft, Nathalie S.M. Lak, Zeinab van Gestel-Fadaie, Marry M. van den Heuvel-Eibrink, Godelieve A.M. Tytgat. A novel ddPCR detects hypermethylated RASSF1A in plasma of patients with pediatric Wilms tumor [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr A026.

OGC O01 - Stomach Cancer Elective Surgery Morbidity and Mortality at 90-Days (HOLD Study): A Prospective, International Collaborative Cohort Study

Abstract Background Data on multinational 90-day mortality and morbidity rates after surgery for gastric cancer is limited in the literature. This study aimed to understand the 90-day mortality and morbidity outcomes according to GASTRODATA Registry for elective gastric cancer surgery patients and identify risk factors. Method We conducted an international prospective study on ≥18 years patients undergoing elective surgery for gastric cancer with curative intent from 01 April 2022 to 30 September 2022. Known metastatic disease, concurrent secondary cancers, gastrointestinal stromal tumour (GIST) and Siewert type I/II oesophagogastric junction malignancies were excluded. Univariate and multivariate logistic regression was used to identify variables associated to 90-day outcome. Results 380 collaborators from 47 countries submitted data on 1538 patients. Mean age was 64.2 years and 58.5% were males. 90-day morbidity rate was 38.2% (n=587) and mortality rate was 2.9% (n=45). Pre-operative higher CCI or ASA score, pre-operative weight loss >10%, and surgical determinants such as type of gastric resection, positive specimen margin, number of harvested lymph nodes, longer surgery duration and post operative pathological IV staging (p value<0.05) were identified as predictors of postoperative severe complications and mortality. Conclusion Elective gastric cancer surgery has a 90-day morbidity of 38.2% and 90-day mortality of 2.9%, globally. This study identified several factors associated with higher morbidity and exemplified the importance of a unified language on surgical morbidity, pre-habilitation and ongoing audits to enhance patient outcomes.

HPB SO07 - Update to the role of staging laparoscopies in stratifying patients by operability in perihilar cholangiocarcinoma

Abstract Background Radical resection of perihilar cholangiocarcinoma (pCCA) is only an option for the minority presenting at an early stage. Sensitivity of radiological imaging to detect liver and peritoneal metastases is insufficient. Our published 2017 cohort study demonstrated over a quarter of patients with no radiologically detectable metastatic disease were subsequently found to have unresectable disease by staging laparoscopy. However, less than 3% of these patients had preoperative PET-CT, which is now strongly recommended in new national guidance.Advances in CT and MRI sensitivity and increased PET-CT use demanded a reassessment of the need for staging laparoscopy in diagnostic assessment of resectability. Method All patients referred to a tertiary UK hepatobiliary centre with a new diagnosis of radiologically suspected or biopsy-proven pCCA from January 2020 to January 2024 were included (n=305). Patients were identified through a prospectively maintained cancer database. Electronic health records were interrogated for demographic data, preoperative radiological imaging, and details related to operative procedures. Those referred with, or subsequently found to have, any of the following conditions were excluded: recurrent pCCA, intrahepatic or distal cholangiocarcinoma, gallbladder cancer or benign hepatobiliary disease. Results All patients underwent CT chest, abdomen and pelvis. 194 (64%) received contrast MRI liver or MRCP. Based on these, 242 (79%) had locally advanced or metastatic cancer, comorbidities precluding resection or otherwise declined surgery. PET-CT was employed to investigate all remaining 63 patients, which discovered metastatic disease in 16 patients (25%). 47 patients proceeded to staging laparoscopy. This yielded metastatic or locally advanced disease in five patients (11%); a near-statistically significant drop compared to our previously published yield of 27.2% (p=0.060). Two (5%) had open-and-close laparotomies for irresectable disease, compared to 16 of 114 (14%) in our 2017 publication (p=0.14). Conclusion Technological advances of CT and MRI and use of PET-CT have vastly improved detection of irresectable pCCA. This study reinforces the merits of PET-CT in identifying occult disease, and we would encourage units that undertake these resections to utilise this imaging modality prior to undertaking staging laparoscopy. Nevertheless, peritoneal spread remains challenging to exclude radiologically, and diagnostic laparoscopy endures as a vital tool in the prevention of open-and-close laparotomies.

Abstract B039: Clinical liquid biopsy testing for detecting actionable genomic alterations in children and young adults with advanced solid tumors

Abstract Introduction: Given the paucity of data regarding circulating tumor DNA (ctDNA) as a modality to guide diagnosis and treatment for children with solid tumors, we conducted a prospective study to determine the frequency of diagnostic and targetable genomic variants identified by clinical ctDNA testing. Methods: GAIN is a prospective, multi-center molecular profiling study that was amended to include clinical ctDNA testing starting in January 2022. Patients were eligible if they were <30 years old with a refractory, recurrent, newly diagnosed with 2-year EFS <50%, or rare solid tumor and had measurable or evaluable disease. Paired tumor and blood samples underwent next generation sequencing (NGS) in CLIA-certified laboratories, with return of results to clinicians. Serial ctDNA samples were analyzed by F1LCDx (FoundationOne® Liquid Companion Diagnostic), an FDA-approved clinical ctDNA test that includes hybrid capture sequencing of a 300+ gene panel, comprehensive multi-omics ctDNA tumor fraction (TF) assessment, tumor mutational burden, and microsatellite instability status. We analyzed comprehensive NGS tissue-based and F1LCDx sequencing data for each participant to compare findings from each sample. The detection of alterations was stratified by ctDNA burden as assessed by ctDNA TF. We calculated the prevalence of ctDNA in our cohort, overall and by disease type. Chart review provided additional patient characteristic, diagnostic, therapeutic, and disease assessment data. The clinical impact of ctDNA results was qualitatively described. Results: The cohort included 35 patients, with a total of 70 F1LCDx samples (range 1-6 samples per patient). There were 16 unique tumor types represented, most commonly osteosarcoma (n = 8), neuroblastoma (n = 8), and Ewing sarcoma (n = 4). 77% of patients had metastatic disease at time of first ctDNA sample collection. 51% of patients had detectable ctDNA TF in at least one sample. Overall, 60% of patients had at least one actionable genomic alteration (range 0-14 variants per patient) identified in liquid biopsy samples, including 3 patients with actionable variants identified with undetectable ctDNA TF. We observed case vignettes in which ctDNA results informed clinical decisions, including disease assessment and treatment guidance. An illustrative example was the identification of ALK variants in relapse blood samples of two patients with neuroblastoma that were not identified in baseline tissue, which were targeted in both instances. Data collection and analyses as described in the methods are ongoing. Conclusion: In this prospective cohort of children and young adults with advanced solid tumors, ctDNA was detectable across a wide range of diagnoses and 60% of patients had at least one diagnostic or potentially targetable alteration identified. These data demonstrate that liquid biopsy yields actionable genomic variants, particularly when ctDNA burden is elevated. We will determine the association of these variants with those identified in tissue samples and their clinical implications in ongoing analyses. Citation Format: Holly J Roberts, Hannah Comeau, Steven G Dubois, Katerina Hoskova, Douglas I Lin, Brian D Crompton, Katherine A Janeway, David S Shulman. Clinical liquid biopsy testing for detecting actionable genomic alterations in children and young adults with advanced solid tumors [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr B039.

Foreign body giant cell reaction due to Durolane (hyaluronic acid derivative) injection - A case report.

Osteoarthritis can be treated with hyaluronic acid derivatives, such as Durolane manufactured by Bioventus (Hill, 2017), particularly when conventional treatments prove ineffective. In osteoarthritic joints, the synovial fluid has a lower concentration of hyaluronic acid compared to healthy joints. Intra-articular therapy with exogenous hyaluronic acid can improve viscoelastic properties, enhance chondrocyte synthesis, reduce cartilage degradation, and alleviate pain associated with osteoarthritis (Migliore and Procopio in Clin Cases Miner Bone Metab. 12:31-3, 2015). This report describes a unique case where a patient, following Durolane administration, presented with an intraosseous foreign body giant cell reaction. A 2.5-cm lesion was identified on MRI, initially raising concerns for metastatic disease or multiple myeloma. Subsequent IR CT-guided biopsy confirmed a diagnosis of "foreign body giant cell reaction to gel," linked to the Durolane injection. This case presents a contribution to the literature due to the scarcity of documented instances where Durolane injection leads to an intraosseous foreign body giant cell reaction resembling metastatic disease. While a solitary intraosseous pseudotumor, reportedly due to viscosupplement injection, has been published, it differs in both location (hip vs. knee) and imaging characteristics from the presented case. The manifestation of a foreign body giant cell reaction after Durolane injection, mimicking malignant osseous lesions, merits attention among practitioners. This case underscores the necessity for further research into potential adverse reactions associated with hyaluronic acid derivatives.

Prognostic factors in clear cell sarcoma: an analysis of soft tissue sarcoma in 43 cases.

Clear cell sarcoma (CCS) of tendons and aponeuroses and CCS-like malignant gastrointestinal neuroectodermal tumor/sarcoma (GINET) are characterized by frequent local and distant relapses, alongside with low efficacy of all systemic treatments. We aimed to collect a comprehensive dataset to identify prognostic factors and treatment outcomes. We performed a retrospective single center analysis for diagnosed CCS and GINET on demographic, tumor, treatment and survival data. We identified 43 patients (w:25, m:18) with a median follow-up of 35mo and a 5y-OS-rate of 42%. At diagnosis the median age was 42yrs. Median tumor size was 3.6cm (0.3-11.1cm), and 24/26 (94%) tissues analyzed at our institute were EWSR1::ATF1-translocation-positive. Distant extremities (incl. knee or elbow) were affected in 72.5%. Of note, 79.5% received an excisional biopsy (benign histology suspected in 30.2%) leading to frequent incomplete resection. Final R0 status correlated significantly (p = 0.017) with longer survival rates compared to R + status in localized CCS (N0M0, 5-yr OS 0% vs 64%). Radiation and systemic treatment had limited antitumor effects while isolated limb perfusion was active in some patients. 18.6% of patients showed lymphatic spread and 20.9% distant metastases. Presence of initial M + was associated with a dismal survival of 1.4years (M +) vs 7.1years (M0; p < .001). We here present one of the largest clinical cohorts of patients with CCS/GINET. Our data underscores the exceptional risk of metastatic disease even in small tumors. As systemic treatment and radiation showed limited efficacy, complete resection was the most important treatment option.

Abstract A005: Microbind affinity blood filter platform technology with affinity for heparan sulfate rapidly removes circulating epithelial tumor and cancer stem cells while downregulating VEGF A in patients with advanced and refractory solid tumors

Abstract Background: Vascular endothelial-derived growth factor A(VEGFA) has been proven to be integral in the molecular pathogenesis of metastasis and tumor growth. It is known that VEGF is responsible for encoding a heparin-binding protein, which exists as a disulfide-linked homodimer which serves to promote the propagation and movement of vascular endothelial cells. This is essential for both pathological and physiological angiogenesis. CETC and cancer stem cells are potent secretors of VEGFA which is an essential cytokine that facilitates new vascular vessel formation, thus further enabling metastatic tumor growth. Methods: We developed a new extracorporeal microbind affinity blood filter platform technology with active heparane sulfate receptors (onco seraph). Heparin is cleaved into short chains which are permanently covalently bound to ultrahigh molecular weight polyethylene (UHMWPE) surfaces to create a new composition of matter. Liquid biopsy established high capacity for the rapid removal of CETC and PDAC cancer stem cells. Onco seraph was investigated in a dose escalation phase I/II clinical trial in adult patients with advanced and relapsed/refractory solid (what) and/or PDAC after standard of care and other lines of therapy failed in preventing disease progression. Results: To date 12 patients have been treated with onco seraph. No patient experienced treatment related adverse effects. Best responses include complete responses and stable disease at a low oncoseraph dose. Greatest efficacy was observed for patients whose liquid biopsy revealed significant and rapid reduction of CETC and cancer stem cells, followed by greater than 50% reduction in circulating VEGFA. Conclusions: This data supports the conclusion that the onco seraph platform technology is safe and has a high therapeutic value as an extracorporeal treatment for metastatic solid tumors refractory to standard of care. Citation Format: Sanja ILIC, Vedran Premuzic, Robert Ward. Microbind affinity blood filter platform technology with affinity for heparan sulfate rapidly removes circulating epithelial tumor and cancer stem cells while downregulating VEGF A in patients with advanced and refractory solid tumors [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr A005.

Endocervical Adenocarcinoma With Micropapillary Component, a Distinct Histological Subtype Associated With Aggressive Behavior and Poor Prognosis: A Clinicopathologic Study of 10 Patients.

Objective. Endocervical adenocarcinoma (ECA) with a micropapillary component is considered to be associated with aggressive behavior and poor prognosis. So far, only limited studies investigated this histological subtype of ECA in the literature. In this study, we present a clinicopathological analysis of 10 such tumors. Methods. We retrieved ten ECAs with a micropapillary component between January 2014 and July 2023 and analyzed their clinicopathologic features. Results. At diagnosis, nine tumors (90%) were of advanced clinical stage (FIGO stage ≥ IIA), nine tumors (90%) exhibited deep stromal invasion (≥2/3 of the cervix), and three tumors (30%) showed parametrial involvement. Lymphovascular invasion was evident in all tumors (100%), and lymph node metastasis was found in eight tumors (80%). Among the 10 patients, six were alive without disease (60%), one had a recurrent/later metastatic tumor (10%), and three died from the disease (30%). Furthermore, eight tumors were positive for PD-L1 expression (80%), while only one tumor showed HER2 overexpression (10%), and one tumor exhibited p53 mutant-type staining (10%). Conclusion. Endocervical adenocarcinomas with micropapillary components are associated with aggressive clinical behavior and a poor prognosis, which can be found in various conventional histological types of ECAs regardless of the HPV status. The high prevalence of PD-L1 expression suggests that patients with micropapillary ECAs may be good candidates for PD-L1 inhibitor treatment.

Abstract A011: Identifying surface biomarkers from colorectal cancer-derived extracellular vesicles using a bead-based approach

Abstract Colorectal cancer (CRC) accounts for the second most common cause of cancer-related mortality globally. The search for prognostic biomarkers subserving to detected cancer at an early stage, to improve tumor classification and to stratify therapeutic patient management groups is actively ongoing. Surgical resections and chemotherapeutics remain a mainstay in the cure and control of solid tumors. Despite their usefulness in the clinical scenario, shortcomings are well recognized, leading researchers to investigate for a circulating source of tumor-derived materials in biofluids, including mostly circulating tumor DNA (ctDNA), circulating tumor cells (CTCs) and extracellular vesicles (EVs). Characterization of EVs surface markers and assessment of their molecular cargo, can lead to the identification of early metastatic colorectal cancer signatures. Cell surface markers of three CRC cell lines (HCT116, DLD1 and LOVO) were characterized using flow cytometry. EVs were isolated from 48-hour serum-deprived cell lines using the polymer pull down method (20% Polyethelene glycol (PEG)). The concentration and particle size distribution of isolated EVs was checked by the NanoSight NS300. Avidin-coated Polystyrene beads were conjugated with CD9 biotin antibodies; and were used to immunoprecipitated (IP) CD9+ EVs. Following a 2-hour incubation and a series of washing steps, the PE-labelled antibodies of interest (CD9, CD63, EpCAM, Vim, CD44, CD133 and CD29) were added and read using xMAP technology. To increase the number of surface markers that can be read in tandem, IP was multiplexed using EpCAM, CD9 and CD63. This was followed by the isolation of EVs from plasma of metastatic CRC patients (N=3) using size exclusion chromatography (Izon qEV1 columns-35nm in conjunction with the automatic fraction collector) and polymer pull down. The concentration and size were assessed by Nanosight NS300 and surface markers of tumor-derived EVs were assessed using the immunoprecipitation technique on the Luminex system. Comparison of signals between EVs isolated from cell lines and matched originator cells showed that cell surface signatures in EVs match closely with their originator cells. The mean fluorescence intensities quantified cell surface marker expression in the different exosome-bead complexes. Interestingly, one of the metastatic cancer patients with invasive colorectal adenocarcinoma and lung metastasis showed EpCAM and Vimentin positivity suggesting epithelial-mesenchymal plasticity. These results show the importance of multiplexing both at the level of immunoprecipitation and at end-measurement, given the dynamics of cell surface marker changes during disease progression. The use of the mesenchymal marker; Vimentin in a panel for cell surface markers stems from studies on biotinylated cell surface markers on various cancer cell lines and identification of Vimentin in such models. Interestingly we find Vimentin on EVs surface in this study. Citation Format: Jessica Debattista, Laura Grech, Christian Scerri, Malcolm Buhagiar, Godfrey Grech. Identifying surface biomarkers from colorectal cancer-derived extracellular vesicles using a bead-based approach [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr A011.

Abstract A017: High-throughput microfluidic enrichment of circulating tumor cells from entire diagnostic leukapheresis for comprehensive liquid biopsy

Abstract Circulating Tumor Cells (CTCs) in blood containing a complete set of single-cell analytes, including high-molecular-weight DNA, intact RNA, and both intracellular and cell-surface proteins serve as a promising tool in cancer diagnosis and prognosis through liquid biopsy. However, their clinical utility has been limited by their low blood concentration and inadequate enrichment technology. Diagnostic leukapheresis holds promise for isolating sufficient CTCs, but enriching ultra-rare cells from large, complex fluid volumes remains challenging. To address this, we developed the LPCTC-iChip, a high-throughput microfluidic device that uses high-flow channels and force-amplifying magnetic lenses to deplete hematopoietic cells, allowing tumor epitope-agnostic screening of massive blood volumes. Here, we present the first application of LPCTC-iChip to entire diagnostic leukapheresis products (leukopak) from seven patients with metastatic cancer, processing an average of 5.83 liters of blood per patient. Enumeration and morphology characterization of CTCs using multispectral immunofluorescence imaging identified high CTC yields - a mean of 10,057 CTCs per leukopak and showed notable intra- patient heterogeneity. Nuclear and cell diameter of CTC varies within individual patients, with 67% overlapping in size with patient-matched white blood cells. Paired single-cell DNA and RNA sequencing uncovered subclonal aneuploidy and distinct signaling pathways within CTCs with shared, clonal copy number variation (CNV) patterns. A subset of small aneuploid cells lacking epithelial markers was enriched for neuroendocrine characteristics in prostate cancer samples. The mutations found by FDA-approved genetic tests in either biopsied metastatic lesions or blood-based ctDNA sampling were reproduced by whole exome sequencing of a pool of CNV-confirmed CTCs. In addition, the sequencing revealed additional cancer-related high allele frequency variants that are not detectable with ctDNA or tissue needle biopsies. Overall, this high-throughput CTC enrichment from complete leukapheresis combined with multispectral imaging and single cell sequencing technology enables a comprehensive, cell-based liquid biopsy approach for early cancer detection and therapeutic responses monitoring. Citation Format: Avanish Mishra, Shih-Bo Huang, Taronish Dubash, Risa Burr, Jon F. Edd, Ben S. Wittner, Quinn E. Cunneely, Victor R. Putaturo, Akansha Deshpande, Ezgi Antmen, Kaustav A. Gopinathan, Keisuke Otani, Yoshiyuki Miyazawa, Ji Eun Kwak, Sara Y. Guay, Justin P. Kelly, John Walsh, Linda T. Nieman, Isabella Galler, PuiYee Chan, Michael S. Lawrence, Ryan J. Sullivan, Aditya Bardia, Douglas S. Micalizzi, Lecia V. Sequist, Richard J. Lee, Joseph W. Franses, David T. Ting, Patricia A. R. Brunker, Shyamala Maheswaran, David T. Miyamoto, Daniel A. Haber, Mehmet Toner. High-throughput microfluidic enrichment of circulating tumor cells from entire diagnostic leukapheresis for comprehensive liquid biopsy [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr A017.

Prognostic significance and risk factors of mediastinal lymph node metastasis in esophagogastric junction cancer: a single-center, retrospective study.

Although the optimal extent of lymph node dissection in esophagogastric junction cancer (EGJC) has been reported, the efficacy of mediastinal lymph node dissection remains unclear. We aimed to identify risk factors for mediastinal lymph node metastasis and its prognostic impact in patients with EGJC. A total of 100 consecutive patients who underwent curative surgery for EGJC were eligible. We examined the rates of metastasis, prognosis, and therapeutic value index at each mediastinal lymph node station. In addition, multivariate analyses were performed to identify risk factors for mediastinal lymph node metastasis. The rates of upper, middle, and lower mediastinal lymph node metastases were 12.0%, 20.7%, and 13.2%, respectively. The 5-year overall survival rate was lower in patients with mediastinal lymph node metastasis than in those without mediastinal lymph node metastasis (11.1% vs. 59.2%, p < 0.01). The therapeutic value index was 0 in patients with upper/middle mediastinal lymph node metastasis, and mediastinal lymph node metastasis was an independent prognostic factor (hazard ratio 6.59, 95% confidence interval [CI] 2.48-17.9, p < 0.01). Additionally, the length of esophageal invasion and the presence of hiatal hernia were independent predictors of mediastinal lymph node metastasis (odds ratio 8.21, 95%CI 1.44-46.8, p = 0.02 and odds ratio 7.13, 95%CI 1.22-41.8, p = 0.03). No survival benefit of mediastinal lymph node dissection was observed. Intensive multidisciplinary treatment could be considered in patients with predicted mediastinal lymph node metastasis, such as those with longer esophageal invasion and those with hiatal hernia.

Metabolism, mitochondria and metastasis in kidney cancer.

Metabolism, mitochondria and metastasis in kidney cancer.

A Computational Approach to Characterize the Protein S-Mer Tyrosine Kinase (PROS1-MERTK) Protein-Protein Interaction Dynamics.

Protein S (PROS1) has recently been identified as a ligand for the TAM receptor MERTK, influencing immune response and cell survival. The PROS1-MERTK interaction plays a role in cancer progression, promoting immune evasion and metastasis in multiple cancers by fostering a tumor-supportive microenvironment. Despite its importance, limited structural insights into this interaction underscore the need for computational studies to explore their binding dynamics, potentially guiding targeted therapies. In this study, we investigated the PROS1-MERTK interaction using advanced computational analyses to support immunotherapy research. High-resolution structural models from ColabFold, an AlphaFold2 adaptation, provided a baseline structure, allowing us to examine the PROS1-MERTK interface with ChimeraX and map residue interactions through Van der Waals criteria. Molecular dynamics (MD) simulations were conducted in GROMACS over 100 ns to assess stability and conformational changes using RMSD, RMSF, and radius of gyration (Rg). The PROS1-MERTK interface was predicted to contain a heterogeneous mix of amino acid contacts, with lysine and leucine as frequent participants. MD simulations demonstrated prominent early structural shifts, stabilizing after approximately 50 ns with small conformational shifts occurring as the simulation completed. In addition, there are various regions in each protein that are predicted to have greater conformational fluctuations as compared to others, which may represent attractive areas to target to halt the progression of the interaction. These insights deepen our understanding of the PROS1-MERTK interaction role in immune modulation and tumor progression, unveiling potential targets for cancer immunotherapy.

Metastasis of colon cancer requires Dickkopf-2 to generate cancer cells with Paneth cell properties.

Metastasis is the leading cause of cancer-related mortality. Paneth cells provide stem cell niche factors in homeostatic conditions, but the underlying mechanisms of cancer stem cell niche development are unclear. Here, we report that Dickkopf-2 (DKK2) is essential for the generation of cancer cells with Paneth cell properties during colon cancer metastasis. Splenic injection of Dkk2 knockout (KO) cancer organoids into C57BL/6 mice resulted in a significant reduction of liver metastases. Transcriptome analysis showed reduction of Paneth cell markers such as lysozymes in KO organoids. Single-cell RNA sequencing analyses of murine metastasized colon cancer cells and patient samples identified the presence of lysozyme positive cells with Paneth cell properties including enhanced glycolysis. Further analyses of transcriptome and chromatin accessibility suggested hepatocyte nuclear factor 4 alpha (HNF4A) as a downstream target of DKK2. Chromatin immunoprecipitation followed by sequencing analysis revealed that HNF4A binds to the promoter region of Sox9, a well-known transcription factor for Paneth cell differentiation. In the liver metastatic foci, DKK2 knockout rescued HNF4A protein levels followed by reduction of lysozyme positive cancer cells. Taken together, DKK2-mediated reduction of HNF4A protein promotes the generation of lysozyme positive cancer cells with Paneth cell properties in the metastasized colon cancers.