bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract PO-003: Mitotic enrichment as an efficient radiosensitization strategy
Authors: de Gooijer Mark C. , Slangen Paul L.G. , Çolakoğlu Hilal , Çitirikkaya Ceren H. , El Ouazani Amal , Shah Ronak , Borst Gerben R. , van Tellingen Olaf
Published: April 2021

Radiotherapy remains one of the most effective modalities for anticancer treatment. Boosting the efficacy of radiotherapy is therefore a logical avenue to improve patient survival. We have developed a radiosensitization strategy called ‘induction of mitotic enrichment’. It has long been known that the radiosensitivity of a cell depends to a large extent on the phase of the cell cycle and that especially mitotic cells are especially vulnerable. Enriching the tumor for mitotic cells by arresting them during division prior to each radiotherapy fraction should therefore render the tumor population more sensitive to irradiation. Ideally, induction of mitotic enrichment should be reversible and non-cytotoxic to prevent healthy tissue toxicity and be compatible with clinically applied fractionated radiotherapy regimens. We have now identified an orally available targeted tubulin polymerization inhibitor that can achieve repeated and reversible mitotic enrichment for up to 10 hours prior to radiotherapy, without causing cytotoxicity in vitro or healthy tissue toxicity in vivo. Most importantly, this tubulin inhibitor efficiently radiosensitizes a range of preclinical glioblastoma models in vitro and in vivo and significantly improves survival, but only in a mitotic enrichment setup when given several hours prior to radiotherapy to allow accumulation in mitosis. We have initiated development of mitotic enrichment for GBM, the most common primary malignant brain tumor. Their location and highly aggressive nature renders GBM among the most deadly and devastating of human malignancies. Despite extensive treatment involving surgery and adjuvant chemo-radiotherapy, prognosis is still dismal and novel treatment strategies are urgently needed. Of all available adjuvant therapies, radiotherapy contributes most to extending overall survival. Increasing the efficacy of existing radiotherapeutic regimens therefore offers a logical rationale to improve the survival of GBM patients. We are currently also expanding our preclinical development of mitotic enrichment as a radiosensitization strategy to other mitotic targets and different cancers for which radiotherapy is a mainstay treatment and have thus far achieved promising results in vitro. In parallel, we are preparing a phase 0 trial to demonstrate induction of mitotic enrichment in human GBM.

Citation Format: Mark C. de Gooijer, Paul L.G. Slangen, Hilal Çolakoğlu, Ceren H. Çitirikkaya, Amal El Ouazani, Ronak Shah, Gerben R. Borst, Olaf van Tellingen. Mitotic enrichment as an efficient radiosensitization strategy [abstract]. In: Proceedings of the AACR Virtual Special Conference on Radiation Science and Medicine; 2021 Mar 2-3. Philadelphia (PA): AACR; Clin Cancer Res 2021;27(8_Suppl):Abstract nr PO-003.
Abstract PO-019: Radiotherapy in cancer is associated with a deletion signature that contributes to poor patient outcomes
Authors: Kocakavuk Emre , Anderson Kevin J. , Varn Frederick S. , Johnson Kevin C. , Amin Samirkumar B. , Sulman Erik. P. , Lolkema Martijn , Barthel Floris P. , Verhaak Roel G.W.
Published: April 2021

Diffuse gliomas are highly aggressive brain tumors that invariably relapse despite treatment with chemo- and radiotherapy. Treatment with alkylating chemotherapy can drive tumors to develop a hypermutator phenotype. In contrast, the genomic effects of radiation therapy (RT) remain unknown. We analyzed the mutational spectra following treatment with ionizing radiation in sequencing data from 190 paired primary-recurrent gliomas from the Glioma Longitudinal Analysis (GLASS) dataset and 3693 post-treatment metastatic tumors from the Hartwig Medical Foundation (HMF). We identified a significant increase in the burden of small deletions following radiation therapy that was independent of other factors. These novel deletions demonstrated distinct characteristics when compared to pre-existing deletions present prior to RT-treatment and deletions in RT-untreated tumors. Radiation therapy-acquired deletions were characterized by a larger deletion size (GLASS and metastatic cohort, P=1.2e-02 and P=8e-11, respectively; Mann-Whitney U test), an increased distance to repetitive DNA elements (P<2.2e-16, Kolmogorov-Smirnov test) and a reduction in microhomology at breakpoints (P=3.2e-02, paired Wilcoxon signed-rank test). These observations suggested that canonical non-homologous end joining (c-NHEJ) was the preferred pathway for DNA double strand break repair of RT-induced DNA damage. Furthermore, radiotherapy resulted in frequent chromosomal deletions and significantly increased frequencies of CDKN2A homozygous deletions. Finally, a high burden of RT-associated deletions was associated with worse clinical outcomes (GLASS and metastatic cohort, P < 1e-04 and P = 2.6e-02, respectively; Wald test). Our results suggest that effective repair of RT-induced DNA damage is detrimental to patient survival and that inhibiting c-NHEJ may be a viable strategy for improving the cancer-killing effect of radiotherapy. Taken together, the identified genomic scars as a result of radiation therapy reflect a more aggressive tumor with increased levels of resistance to follow up treatments.

Citation Format: Emre Kocakavuk, Kevin J. Anderson, Frederick S. Varn, Kevin C. Johnson, Samirkumar B. Amin, Erik. P. Sulman, Martijn Lolkema, Floris P. Barthel, Roel G.W. Verhaak. Radiotherapy in cancer is associated with a deletion signature that contributes to poor patient outcomes [abstract]. In: Proceedings of the AACR Virtual Special Conference on Radiation Science and Medicine; 2021 Mar 2-3. Philadelphia (PA): AACR; Clin Cancer Res 2021;27(8_Suppl):Abstract nr PO-019.
Abstract PO-035: Understanding the role of hypoxia and PTEN loss in driving prostate cancer progression
Authors: Suvac Alexandru , Rebello Richard , Lyons Stephen , Bristow Robert G.
Published: April 2021

Prostate cancer is the most commonly diagnosed cancer in the UK. Increased levels of prostate hypoxia have been shown to drive tumor aggression as an adverse prognostic factor following surgery or radiotherapy, especially when combined with tumor genetic instability. Our recent collaborative bioinformatic studies (Bhandari et al, Nat. Genetics 2019; Nat. Communications 2020) have shown various copy number changes to be enriched in hypoxic tumors, with mono- or bi-allelic loss of the PTEN being the most significant. To delineate a direct relationship between hypoxia and PTEN loss, we interrogated the impact of chronic hypoxia (1% O2, 0.2% O2; 72 h) on PTEN oncogenic signaling and asked whether this translates to improved fitness and survival in non-transformed and transformed PTEN isogenic cell lines. Despite increased oncogenic signaling (increased phospho-AKT) of PTEN-null 22Rv1 cells and hTERT-immortalized prostate epithelial cells (GHM) under chronic hypoxic gassing conditions, this did not lead to increased adaption and tumor clone survival. Differential survival for the PTEN isogenic cell lines following prostate cancer therapies (i.e. IR, PARPi, cDDP, taxols) under oxia and hypoxia are underway to further this work and determine a potential translational importance to clinical treatment. Independent evidence also points to hypoxia/PTEN loss in deregulating centrosome biology as a potential mechanism for genetic instability under hypoxia. Hence, we also hypothesized that the relationship between PTEN and hypoxia might be unearthed if aberrant unstable mitoses following deregulation of spindle poles under hypoxia selectively adapt and have a survival advantage. Quantitative immunofluorescence of centrosome aberrations, using antibodies to phospho-Histone H3 and Centrin-1, show significant increases in centriole numbers (p < 0.05) in mitotic cells under chronic hypoxic conditions and an increase in the incidence of micronuclei (p < 0.005), specifically in chronic hypoxic cells with PTEN loss, supporting our hypothesis. Current studies are investigating in situ staining of centrosomes as a readout of chromosomal instability (CIN) in primary prostate cancer tissues with variable PTEN status as a new biomarker for aggressiveness to link hypoxia-associated genetic instability with novel clinical trial development. (Funded by Cancer Research UK as a core grant to RGB).

Citation Format: Alexandru Suvac, Richard Rebello, Stephen Lyons, Robert G. Bristow. Understanding the role of hypoxia and PTEN loss in driving prostate cancer progression [abstract]. In: Proceedings of the AACR Virtual Special Conference on Radiation Science and Medicine; 2021 Mar 2-3. Philadelphia (PA): AACR; Clin Cancer Res 2021;27(8_Suppl):Abstract nr PO-035.
Abstract PO-053: Targeting focal adhesions to limit radiation-induced glioblastoma cell invasion
Authors: Decotret Lisa R. , Shi Rocky , Bennewith Kevin L.
Published: April 2021

Introduction: Glioblastoma multiforme (GBM) is a grade IV brain tumor that is considered incurable with a median overall survival of 15 months. This low survival rate is partially attributed to the highly infiltrative nature of GBM cells. A key component of GBM cell invasion is the degradation of extracellular matrix (ECM), which is facilitated by the formation of invadosomes: focal adhesions (FAs) and invadopodia. The majority of GBM patients will receive a conventional radiation schedule of 2 Gy in 30 fractions, while a subset of patients will receive a hypofractioned radiation schedule of ~3 Gy in 15 fractions. Recent pre-clinical reports indicate the population of GBM cells that survive ionizing radiation (IR) exhibit increased invasiveness, suggesting IR may regulate FA and invadopodia function. Understanding how IR affects invadosome biology may lead to new therapeutic strategies to halt the invasion of GBM cells and increase overall survival of patients treated with radiation. Results: We found that irradiated GBM cells were significantly more invasive in vitro. GBM cells irradiated with 0-4 Gy and plated onto fluorescently-tagged gelatin to analyze ECM degradation. Cells exposed to 0-2 Gy IR produced small dot-like patterns of degradation resembling active invadopodia. However, cells exposed to 3-4 Gy IR produced oblong-shaped patterns of degradation at the periphery of the cell resembling active FAs. Using immunofluorescent markers of invadopodia (TKS5/actin) and FAs (vinculin), we found total focal adhesion and invadopodia formation remained unchanged. Rather, we found that exposing cells to 3 Gy IR increased pFAK-397 in FA structures suggesting IR increases ECM degradation by promoting FA activity. Conclusions: These data suggest higher IR doses promote FA-mediated ECM degradation and GBM cell invasion. We are currently investigating the impact of IR on FAs, ECM degradation, and GBM cell invasion in murine brain slices ex vivo.

Citation Format: Lisa R. Decotret, Rocky Shi, Kevin L. Bennewith. Targeting focal adhesions to limit radiation-induced glioblastoma cell invasion [abstract]. In: Proceedings of the AACR Virtual Special Conference on Radiation Science and Medicine; 2021 Mar 2-3. Philadelphia (PA): AACR; Clin Cancer Res 2021;27(8_Suppl):Abstract nr PO-053.
Abstract PO-054: Correlation of hematological toxicity with the bone marrow radiation dose and volume during concurrent chemo radiation in patients with cervical cancer
Authors: Madabhavi Irappa , Sagar Ragavendra , Patel Apurva , Sarkar Malay
Published: April 2021

Aims & Objective: Hematological toxicity is common in patients with cervical cancer treated with concurrent chemo radiotherapy (CT-RT), so the purpose is to assess this hematological toxicity and correlate the toxicity with the dose and volume of bone marrow included in the field of radiation. Materials & Methods: Twenty-five patients with histologically proven cervical cancer attending to our cancer center from July 2008-August 2009 were the subjects of this study. Patients were treated on 6 MV linear accelerator with a radical intent with concurrent chemotherapy using cisplatin 50 mg weekly. The planning CT was done for all the patients before the treatment and contouring of the pelvic bone marrow apart from other organs at risk was done. Hematological toxicity was assessed using RTOG common toxicity criteria weekly during and at 2 weeks after the completion of the treatment. Results: A total of 25 patients on CT-RT treatment were assessed. Sixteen patients were in locally advanced stage. The variation in HB, TLC, Platelets, and ANC counts from the baseline to 2 weeks after chemo radiotherapy were assessed. Grade II anemia was observed in 12 and Grade III in 2 patients. There were no toxicity as far as WBC and platelets were considered. There was also no correlation between the volume of bone marrow included in the field of irradiation and appearance of anemia. Conclusion: CT-RT for cervical cancer is safe and is associated with minimal hematological toxicity in the form of anemia. The toxicity is same for different volumes of bone marrow included in the field of irradiation with both conventional as well as 3DCRT technique. The toxicity observed is probably contributed by Cisplatin.

Citation Format: Irappa Madabhavi, Ragavendra Sagar, Apurva Patel, Malay Sarkar. Correlation of hematological toxicity with the bone marrow radiation dose and volume during concurrent chemo radiation in patients with cervical cancer [abstract]. In: Proceedings of the AACR Virtual Special Conference on Radiation Science and Medicine; 2021 Mar 2-3. Philadelphia (PA): AACR; Clin Cancer Res 2021;27(8_Suppl):Abstract nr PO-054.
Abstract PO-064: Predicting molecular phenotypes with scRNAseq: An assessment of unsupervised machine learning methods
Authors: Dunca Anastasia , Adler Frederick R. , Smithson Mark W.
Published: April 2021

We evaluated the performance of machine learning models in predicting cell state based on single cell RNA sequencing data (scRNAseq) analysis. scRNAseq quantifies the number of mRNA molecules in individual cells providing information on regulatory relationships between genes and the trajectories of distinct cell lineages in development. Different cells may express different genes at different levels which is key to analyzing the molecular phenotypes of cells. We tested nine machine learning models for their ability to distinguish subclonal cell populations in scRNAseq data by making predictions of cell state. Two scRNAseq datasets were examined: one with cells separated by cell phase: S, G1, or G2/M and one with distinct tumor and non-tumor cells. The pipeline consisted of data filtering, dimensionality reduction with Principal Component Analysis, naive projection with Uniform Manifold Approximation and Projection, and cluster analysis using Ward, BIRCH, Gaussian Mixture Model, DBSCAN, Spectral, Affinity Propagation, Average Linkage Agglomerative Clustering, Mean Shift, and K-Means models. These represent centroid, density, hierarchical, distribution, and graph-based clustering. Models failing to generate the true number of cells of each type were eliminated from further evaluation. The rest were evaluated on accuracy on a true cell type versus predicted cell type basis with a mapping algorithm. The true molecular phenotypes of the plotted cells were annotated onto the data points for a biological examination of the clusters. Six models divided G2 vs. S cells; Spectral and Ward clustering performed best with ~60% accuracy on average. Parallel to that result, seven models divided the total population into the tumorous and non-tumorous subclonal cell populations; Ward and BIRCH clustering performed best with ~80% accuracy on average. Overall, unsupervised machine learning models were effective in predicting molecular phenotypes of cells by accurately identifying subclonal cell populations and analyzing the relationship between gene expression levels and cell state. BIRCH, Ward, and Spectral clustering showed the highest performance.

Citation Format: Anastasia Dunca, Frederick R. Adler, Mark W. Smithson. Predicting molecular phenotypes with scRNAseq: An assessment of unsupervised machine learning methods [abstract]. In: Proceedings of the AACR Virtual Special Conference on Radiation Science and Medicine; 2021 Mar 2-3. Philadelphia (PA): AACR; Clin Cancer Res 2021;27(8_Suppl):Abstract nr PO-064.
Abstract PO-077: Study evaluating metastatic castrate resistant prostate cancer (mCRPC) treatment using 177Lu-PNT2002 PSMA therapy after second-line hormonal treatment (SPLASH) - Trial in progress
Authors: Chi Kim N. , Metser Ur , Czernin Johannes , Calais Jeremie , Prasad Vikas , Eiber Matthias , Shore Neal , Jensen Jessica , Fleshner Neil , Tagawa Scott , Sartor Oliver
Published: April 2021

Background: Treatment options with minimal toxicity and novel mechanisms of action are urgently needed to improve clinical outcomes from mCRPC. Prostate-specific membrane antigen (PSMA)-targeted radioligand therapy (RLT) represents a new treatment for patients with PSMA-avid mCRPC. 177Lu-PNT2002 (also known as [Lu-177]-PSMA-I&T) is a PSMA-targeting agent and studies have shown demonstrable promising initial data. This trial seeks to prospectively evaluate the efficacy of 177Lu-PNT2002 for men with progressive mCRPC after androgen receptor axis-targeted (ARAT) therapy. Methods: This is a multi-center, open-label, phase III study. All patients must be at least 18 years of age, have documented progressive mCRPC at time of screening, high PSMA expression by PSMA PET/CT per blinded independent central review (BICR), chemotherapy naïve for CRPC and unfit or unwilling to receive chemotherapy. The study will commence with a 25-patient dosimetry lead-in. In the dosimetry phase, patients will receive up to four cycles of 177Lu-PNT2002 at 6.8 GBq every 8 weeks. In the randomization phase, approximately 390 patients will be randomized in a 2:1 ratio to receive 177Lu-PNT2002 (Arm A) versus enzalutamide or abiraterone (with prednisone or dexamethasone) (Arm B). Patients randomized to Arm B have an option to crossover to 177Lu-PNT2002 treatment after BICR-assessed radiologic progression. The primary endpoint is Radiological progression-free survival (rPFS) assessed by BICR using Response Evaluation Criteria in Solid Tumors (RECIST) 1.1 (soft tissue) and Prostate Cancer Working Group 3 (PCWG3) (bone) criteria. Key secondary endpoints include objective response rate, duration of response, PSA response, and overall survival. The study is powered at 90% to test the alternative hypothesis of a hazard ratio (HR) ≤ 0.66 at an α of 0.025. ClinicalTrials.gov identifier: NCT04647526.

Citation Format: Kim N. Chi, Ur Metser, Johannes Czernin, Jeremie Calais, Vikas Prasad, Matthias Eiber, Neal Shore, Jessica Jensen, Neil Fleshner, Scott Tagawa, Oliver Sartor. Study evaluating metastatic castrate resistant prostate cancer (mCRPC) treatment using 177Lu-PNT2002 PSMA therapy after second-line hormonal treatment (SPLASH) - Trial in progress [abstract]. In: Proceedings of the AACR Virtual Special Conference on Radiation Science and Medicine; 2021 Mar 2-3. Philadelphia (PA): AACR; Clin Cancer Res 2021;27(8_Suppl):Abstract nr PO-077.
Abstract PO-082: Radiation therapy enhances the potential of the oncolytic virus in the treatment of osteosarcoma
Authors: Hossain Elora , Higashino Fumihiro
Published: April 2021

Background: Combination of neoadjuvant chemotherapy and surgery is the standard treatment approach for osteosarcoma. Despite aggressive treatment protocol, ~40% of patients do not respond and relapse, the survival rate is poor for these patients. Conditionally replicative adenovirus (CRAds) destroying cancer cell while sparing normal cells is a new treatment approach for cancers. We developed a new oncolytic adenovirus which grows using mRNA stabilization system found in many cancer cells. Our recent report showed that this virus replicated in cancer cells where AU-rich element (ARE) containing mRNA is stabilized. ARE-mRNA is stabilized in the majority types of cancers mediate by binding with HuR (Human antigen R), which exports target ARE-mRNA to the cytoplasm. Radiotherapy (RT) is not a first-line treatment option for osteosarcoma as osteosarcoma cells are not killed by radiation easily. But in certain conditions radiotherapy can be used where surgical resection is not technically possible without undue morbidity or when surgery is refused. In response to ionizing radiation and UV radiation DNA damage response (DDR) can influence HuR action either by the increasing cytoplasmic HuR abundance or affecting the binding affinity to target ARE-mRNA. The purpose of the present study was to analyze the efficacy of the combination treatment of RT and oncolytic adenovirus for osteosarcoma. Materials and Methods: E4rof6-deleted adenovirus (dl355) and human osteosarcoma cell lines U2OS and Saos2 were used in this study. Cell viability assays were carried out to evaluate the cytotoxicity and synergism of dl355 and RT combination. Chou and Talalay’s median-effect method was used to calculate the combination index. CPE assay was used to assess the cytopathic effect of the combination. Viral replication with or without radiation combination was investigated by hexon staining. Western blot analysis was performed to examine viral protein expression. Results: We found that RT treatment enhanced the cytoplasmic relocalization of HuR; thus irradiation up-regulated the propagations of dl355. Also, radiation-induced upregulation of viral protein expression was observed by western blotting. The RT and dl355 combination treatment increased the cytolysis of U2OS, and Saos2 cells in a synergistic manner compared to RT or virus only. Conclusion: Radiotherapy up-regulated of oncolytic adenovirus, which grows using the ARE-mRNA stabilization system, and this cytolytic effect provides a rationale for this combination. This combination approach has the possibility of a promising strategy for osteosarcoma.

Citation Format: Elora Hossain, Fumihiro Higashino. Radiation therapy enhances the potential of the oncolytic virus in the treatment of osteosarcoma [abstract]. In: Proceedings of the AACR Virtual Special Conference on Radiation Science and Medicine; 2021 Mar 2-3. Philadelphia (PA): AACR; Clin Cancer Res 2021;27(8_Suppl):Abstract nr PO-082.
Abstract PO-095: Intrinsic and acquired resistance to RIG-I induced anti-tumor responses
Authors: Anand Sudarshan , Fraile-Bethencourt Eugenia , Khou Sokchea
Published: April 2021

The immune microenvironment plays a critical role in cancer orchestrating tumor progression and response to therapeutics. Strategies to manipulate the tumor immune microenvironment are invaluable to complement current standard of care treatments. Recently, approaches to target Pattern Recognition Receptors (PRRs) that recognize intracellular and extracellular pathogens have emerged as a promising strategy to enhance immune recognition of tumors. Among the PRRs, intracellular receptors that recognize DNA and RNA in the cells are unique in their ability to respond to both pathogenic and ‘self’ nucleic acids that are generated as a result of aberrant cell division/transcription. We previously showed that targeting a cytosolic DNA sensor three prime exonuclease 1 (TREX1) enhanced immune responses in mouse tumor models including colorectal carcinomas (CRCs). Here we show that a cytosolic RNA sensing pathway, Retinoic acid Induced Gene 1 (RIG-I) acts as a robust immune activator across cancer types by enhancing the cellular and molecular correlates of anti-tumor immune responses. Using a stem loop hairpin RNA structure derived from a H1N1 sequence we elicited a potent RIG-1 response in CRC and breast cancers. Interestingly, activation of RIG also increased expression of TREX1 revealing potential cross-talk between the two pathways. We show that activation of tumor cell RIG-I alone is sufficient to decrease tumor burden with a concomitant broad cellular and molecular anti-tumor response. We observed a group of negative regulators of immune responses, including the NKG2a ligand - HLA-E, mono PARPs and the inhibitory checkpoint protein LAG3, are upregulated in RIG high tumors in cell culture, mouse and human tumors indicating how RIG-I high tumors may develop resistance to immune killing. Combination of RIG-I agonists synergized with inhibition of these resistance mechanisms in a tumor and context dependent manner. To understand intrinsic resistance to RIG signaling, we performed a focused RNAi screen and identified critical positive and negative modulators of RIG-I function in tumor cells. Taken together, our findings illustrate how tumor cell RIG-I is a potent inducer of cellular and molecular immune responses. Our work also highlights tumor cell intrinsic and extrinsic pathways that confer resistance to RIG-I induced anti-tumor responses and provide strategies to mitigate such resistance.

Citation Format: Sudarshan Anand, Eugenia Fraile-Bethencourt, Sokchea Khou. Intrinsic and acquired resistance to RIG-I induced anti-tumor responses [abstract]. In: Proceedings of the AACR Virtual Special Conference on Radiation Science and Medicine; 2021 Mar 2-3. Philadelphia (PA): AACR; Clin Cancer Res 2021;27(8_Suppl):Abstract nr PO-095.
Abstract PO-097: Selective targeting and imaging of orthotopic glioblastoma after a single systemic dose of a novel hydoxyl dendrimer radionuclide
Authors: Sharma Rishi , Sun Minghao , Appiani Santiago , Coelho Richard , McConville Patrick , Alters Susan , Cleland Jeffrey L.
Published: April 2021

Previous studies have demonstrated hydroxyl dendrimers (HDs) are selectively taken up by tumor associated macrophages (TAMs) after a single administration. HDs have also been observed to cross the blood brain barrier in six animal species including dogs and monkeys. The treatment of brain tumors and metastases is hampered by the inability to deliver radiotherapy or cancer agents selectively to the tumor. HDs of two different generations were prepared (HD4 and HD6; both smaller than antibodies) and DOTA was conjugated to the HD through linker arms attached to the hydroxyls on the dendrimer surface. 111In radiolabeling of HD4- and HD6-DOTA resulted in >95% radiochemical purity in both cases and specific activities of 0.621 mCi/mg and 0.638 mCi/mg, respectively. GL-261-luc cells, a syngeneic glioblastoma, were implanted intracranially in 8 female albino C57.bl/6 mice. Tumors were 15-60 mm3 at time of dosing. Mice received a single administration of HD4-DOTA-111In or HD6-DOTA-111In at a dose of approximately 0.5 mg/mouse and 0.3 mCi/mouse. Whole body SPECT/CT imaging was done over time up to 6 days post-dose in 3 animals per compound. The biodistribution and uptake in the brain tumor and other tissues was quantitatively measured using VivoQuant 4.0 (Invicro, Boston). Both HDs rapidly distributed in the systemic circulation after administration. Clearance appeared to be primarily via the kidneys based upon the fraction of injected dose observed in the bladder over time. The % injected dose (%ID)/g in the brain tumor was 2.67 ± 0.45 and 7.05 ± 0.53 at 24 h post-dose for the HD4 and HD6 compounds, respectively. The %ID/g in the contralateral brain section was 0.37 ± 0.17 and 0.90 ± 0.20 at 24 h post-dose for the HD4 and HD6 compounds, respectively. Tumor associated radioactivity persisted for the duration of the study (last time points: HD4, 96 h, 5.27 ± 2.04 %ID/g; HD6, 144 h, 3.20 ± 1.02 %ID/g). These results confirm previous studies in this model demonstrating selective tumor uptake and persistence with 58 fold greater radioactivity at the last time point in brain tumor compared to contralateral brain. The selectivity and prolonged duration of HDs in TAMs will enable the use of the same HD-DOTA constructs to deliver radiotherapeutic 90Y to brain tumors. The HD radiotherapy approach may eliminate the off-target toxicity of current ligand-based approaches.

Citation Format: Rishi Sharma, Minghao Sun, Santiago Appiani, Richard Coelho, Patrick McConville, Susan Alters, Jeffrey L. Cleland. Selective targeting and imaging of orthotopic glioblastoma after a single systemic dose of a novel hydoxyl dendrimer radionuclide [abstract]. In: Proceedings of the AACR Virtual Special Conference on Radiation Science and Medicine; 2021 Mar 2-3. Philadelphia (PA): AACR; Clin Cancer Res 2021;27(8_Suppl):Abstract nr PO-097.
Abstract PR-008: Identification and characterization of the molecular mechanisms of SCLC chemo-radiation resistance
Authors: Carrieri Francesca Anna , Connis Nick , Grasset Eloise , Luidy-Imada Eddie , Ewald Andrew , Marchionni Luigi , Hann Christine , Tran Phuoc T.
Published: April 2021

Small cell lung cancer (SCLC) is among the most aggressive form of lung malignancies and accounts for 15-20% of all lung cancers. It has the tendency to metastasize early, thus limited-stage SCLC patients still receive systemic treatment with chemo-radiotherapy (chemoXRT) for their localized disease. SCLC is exceptionally sensitive to chemoXRT and exhibits high response rates; however, the recurrence rate is almost 100% and patients relapse with tumors that resist further treatments. Elucidating mechanisms of chemoXRT resistance in SCLC is needed to develop improved therapies and positively impact patient outcomes. To better interrogate mechanisms of chemoXRT resistance, we developed a SCLC patient-derived xenograft (PDX) in vivo system for the major molecular subtypes of SCLC (classic and variant). Briefly, PDX tumor bearing mice were treated with: 1) vehicle control; 2) cisplatin plus etoposide (EP); 3) radiotherapy (XRT); and 4) both EP/XRT. A major response was observed within the EP/XRT arm compared to vehicle or single therapy arms. Whole transcriptome profiling among all treatment arms revealed molecular pathways and biological processes associated with chemoXRT resistance. Also, by comparing our data with two previous SCLC patient cohort studies, we identified gene candidates for functional validation of chemoXRT resistance (i.e. ST6GAL1, TNIK and SOHLH2). To enable real-time cellular and molecular analysis of PDX behavior ex vivo and to validate SCLC chemoXRT resistance candidate genes, we established a novel PDX organoid (PDO) model to study the molecular underpinnings of XRT resistance in SCLC. Classic and variant SCLC PDOs still retained the cellular, DNA and RNA markers consistent with their parental PDX molecular subtype classification using array comparative genomic hybridization and RNA-sequencing. We aim to utilize our novel SCLC PDX/PDO models as a tool to identify and validate candidates for chemoXRT resistance to be used as biomarkers and targets to combat chemoXRT resistance in SCLC.

Citation Format: Francesca Anna Carrieri, Nick Connis, Eloise Grasset, Eddie Luidy-Imada, Andrew Ewald, Luigi Marchionni, Christine Hann, Phuoc T. Tran. Identification and characterization of the molecular mechanisms of SCLC chemo-radiation resistance [abstract]. In: Proceedings of the AACR Virtual Special Conference on Radiation Science and Medicine; 2021 Mar 2-3. Philadelphia (PA): AACR; Clin Cancer Res 2021;27(8_Suppl):Abstract nr PR-008.
Early, Postnatal Pulmonary Hypertension Severity Predicts Inpatient Outcomes in Congenital Diaphragmatic Hernia
Authors: Ferguson Dalya Munves , Gupta Vikas S. , Lally Pamela A. , Luco Matias , Tsao KuoJen , Lally Kevin P. , Patel Neil , Harting Matthew T. ,   for The Congenital Diaphragmatic Hernia Study Group
Published: April 2021

Introduction: Pulmonary hypertension (PH) is the major pathophysiologic consequence of congenital diaphragmatic hernia (CDH). We aimed to evaluate the association between early CDH-associated PH (CDH-PH) and inpatient outcomes. Methods: The CDH Study Group registry was queried for infants born 2015–2019 with echocardiograms before 48h of life. PH was categorized using echocardiographic findings: none, mild (right ventricular systolic pressure <2/3 systemic), moderate (between 2/3 systemic and systemic), or severe (supra-systemic). Univariate and multivariate analyses were performed. Adjusted Poisson regression was used to assess the primary composite outcome of mortality or oxygen support at 30 days. Results: Of 1,472 patients, 86.5% had CDH-PH: 13.9% mild (n = 193), 44.4% moderate (n = 631), and 33.2% severe (n = 468). On adjusted analysis, the primary outcome of mortality or oxygen support at 30 days occurred more frequently in infants with moderate (incidence rate ratio [IRR] 1.8, 95% confidence interval [CI], 1.2–2.6) and severe CDH-PH (IRR 2.0, 95% CI, 1.3–2.9). Extracorporeal life support (ECLS) utilization was associated only with severe CDH-PH after adjustment (IRR 1.8, 95% CI, 1.0–3.3). Discussion/Conclusion: Early, postnatal CDH-PH is independently associated with increased risk for mortality or oxygen support at 30 days and utilization of ECLS. Early echocardiogram is a valuable prognostic tool for early, inpatient outcomes in neonates with CDH.
Stapedial Reflex: A Possible Novel Biomarker of Early Bulbar Involvement in Amyotrophic Lateral Sclerosis Patients
Authors: Bombaci Alessandro , Lazzaro Chiara , Bertoli Carola Amalia , Lacilla Michelangelo , Ndrev Drita , Chiò Adriano , Albera Andrea , Calvo Andrea , Canale Andrea
Published: April 2021

Background and Aim: Amyotrophic lateral sclerosis (ALS) is a neuromuscular progressive disorder, characterized by limb and bulbar muscle wasting and weakness. 30% of patients present a bulbar onset, while 70% a spinal outbreak, although most of them develop bulbar impairment later on. Due to the lack of an early biomarker of bulbar involvement, we chose to evaluate the role of stapedial reflex (SR) in order to predict preclinical bulbar impairment in ALS. Materials and Methods: We enrolled 36 ALS patients. We assessed revised-ALS functional-rating-scale and SR for a total of 4 visits. We established the presence of SR, acoustic reflex latency test (ARLT), and SRs Decay. Patients who had not develop bulbar signs at fourth visit continued follow-up up to 15 months. Data were analyzed by using Mann-Whitney U test, Friedman test, and Cox regression analysis. Results: We observed that SRs Decay at 500 and 1,000 Hz is the first parameter of SR to get altered in all ALS patients before the development of bulbar impairment. Twenty-eight patients developed bulbar impairment during the study. We highlighted a correlation between the progression rate of disease and both time of SRs Decay alteration and time of bulbar impairment from disease onset. Four patients who did not develop bulbar impairment had a progression rate lower than the other ones (p < 0.05). Discussion and Conclusions: This study shows that SR Decay test could be a sensitive measure for detecting pre-symptomatic bulbar involvement in ALS and could represent a simple, noninvasive, and useful biomarker of disease progression.
Machine Learning Algorithms for Predicting Fatty Liver Disease
Authors: Pei Xieyi , Deng Qingqing , Liu Zhuo , Yan Xiang , Sun Weiping
Published: April 2021

Background: Fatty liver disease (FLD) has become a rampant condition. It is associated with a high rate of morbidity and mortality in a population. The condition is commonly referred as FLD. Early prediction of FLD would allow patients to take necessary preventive, diagnosis, and treatment. The main objective of this research is to develop a machine learning (ML) model to predict FLD that can help medics to classify individuals at high risk of FLD, make novel diagnosis, management, and prevention for FLD. Methods: Total of 3,419 subjects were recruited with 845 having been screened for FLD. Classification models were used in the detection of the disease. These models include logistic regression (LR), random forest (RF), artificial neural networks (ANNs), k-nearest neighbors (KNNs), extreme gradient boosting (XGBoost), and linear discriminant analysis (LDA). Predictive accuracy was assessed by area under curve (AUC), sensitivity, specificity, positive predictive value, and negative predictive value. Results: We demonstrated that ML models give more accurate predictions, the best accuracy reached to 0.9415 in the XGBoost model. Feature importance analysis not only confirmed some well-known FLD risk factors, but also demonstrated several novel features for predicting the risk of FLD, such as hemoglobin. Conclusion: By implementing the XGBoost model, physicians can efficiently identify FLD in general patients; this would help in prevention, early treatment, and management of FLD.
Inferior Vena Cava Syndrome as a Manifestation of Metastatic Carcinoid Tumor
Authors: Stankard Matthew , Soule Erik , Matteo Jerry
Published: April 2021

Small bowel-origin carcinoid tumor is indolent but may metastasize relentlessly to various sites, including the liver. Over the past 9 years, we have treated a 69-year-old woman who has undergone 5 percutaneous liver ablations, 5 hepatic intra-arterial chemoembolizations, an ovarian cryoablation, and a trans-ventral hernia mesenteric cryoablation. These interventions are all related to her inoperable carcinoid malignancy. After the patient presented with swelling of the abdomen and both lower extremities, computed tomography (CT) angiography was performed, revealing a circumferential hepatic metastatic mass encasing the intrahepatic inferior vena cava (IVC) and extensive third spacing of fluids specific to the IVC distribution below the diaphragm. A venogram of the intrahepatic IVC revealed extrinsic compression causing 95% narrowing of the vessel. A balloon was advanced to the level of the lesion and inflated, increasing the caliber of the vessel. Subsequently, 2 covered aortic stent graft cuffs were deployed in an overlapping fashion within the lumen of the IVC, traversing the area of narrowing. Next, an open-cell aortic dissection stent was placed across both overlapping aortic stents from the renal veins to the hepatic veins. Following this, three 17-gauge cryoablation probes were inserted into the segment 1 intrahepatic lesions encasing the newly stented IVC via an anterior percutaneous approach. Two 10-min freeze cycles were performed with intraoperative CT imaging, demonstrating circumferential coverage of the lesions. Posttreatment venogram revealed patent stent grafts within the intrahepatic IVC, and restoration of vessel patency. No immediate postoperative complications were noted. The patient’s abdominal and lower extremity swelling resolved completely within 1 week after procedure. Two-month follow-up CT demonstrated markedly decreased size of the metastatic lesions and no adverse effects. Six- and 9-month PET-CT scans demonstrated maintained patency of the IVC stent. This palliative procedure allowed the patient to maintain good performance status and alleviated her symptoms of IVC syndrome. The radial force generated by the multiple aortic stents will ostensibly maintain the patency of the intrahepatic IVC. Cryoablation of the encasing metastatic lesion was performed with markedly decreased size of the tumor on the 2-month follow-up.
Long-Term Medical Therapy after Laparoscopic Excision of Ovarian Endometriomas: Can We Reduce and Predict the Risk of Recurrence?
Authors: Del Forno Simona , Cofano Maria , Degli Esposti Eugenia , Manzara Federica , Lenzi Jacopo , Raimondo Diego , Arena Alessandro , Paradisi Roberto , Casadio Paolo , Seracchioli Renato
Published: April 2021

Objectives: Up to 32% of women experience anatomic recurrence after conservative surgery for endometriomas, while pain recurs in 10–40% of cases. Long-term postoperative hormonal therapy is recommended to prevent disease recurrence. We evaluated the efficacy of long-term therapy with estroprogestins (EPs) or progestins (Ps) in preventing endometrioma recurrence, as identifiable cysts and subjective symptoms, after laparoscopic excision. Design: This retrospective cohort study included 375 women submitted to laparoscopic endometrioma excision. Women were followed up at 6 and 12 months and then yearly after surgery. Based on postoperative medical therapy, women were divided into 4 groups: nonusers, cyclic EP users, continuous EP users, and progestogen users. Materials, Setting, Methods: Anamnestic and anthropometric characteristics were collected as well as clinical and surgical data. Gynecological examination, and transvaginal and transabdominal ultrasound scans were performed. Pain (numerical rating score >5) and endometrioma recurrence at ultrasound (ovarian cyst with typical sonographic features ≥10 mm in mean diameter) were recorded at each examination. The reoperation rate in women with recurrence was investigated. Results: The median follow-up was 3.7 years with a maximum of 16.7 years. Most patients used EPs (119 cyclic and 61 continuous users), 95 used P, and 100 were nonusers. In 135 women (36%), endometriotic cyst recurrence was diagnosed, with a mean diameter of 18.7 ± 10.8 mm (range 10–55 mm). The median recurrent cyst-free time was 7.9 years (95% CI 5.8–10.8). Dysmenorrhea was the first symptom to reappear, affecting 162 patients (43.2%). Upon multivariable regression analysis, continuous users had a lower risk of relapse (OR 0.56, 95% CI 0.32–0.99), in terms of both cysts and symptom recurrence, than patients who received no medications. The reoperation rate was 16.2%. Limitations: The main limitation of this study is its retrospective design. Also, women switching therapies throughout the follow-up period were sorted into one of the study groups based on the longest treatment taken, without considering the discontinuation rates. Conclusions: Long-term EPs, administered in a continuous regimen and starting immediately after conservative surgery for endometriomas, seem to reduce the disease recurrence risk.
Medical Thoracoscopy Implementation after a European Respiratory Society Course Held from 2003 to 2016: A Survey
Authors: Froudarakis Marios E. , Anevlavis Stavros , Marquette Charles-Hugo , Astoul Philippe
Published: April 2021

Background: Medical thoracoscopy (MT) is an important procedure in the management of patients with pleural diseases. Objectives: We designed a survey to explore whether the participants of our courses implement MT at their hospital after attending the course as no real-life data exist. Methods: We distributed by e-mail a questionnaire to the participants of the courses. The questionnaire included general information about the participants, the precourse experience on MT, the postcourse implementation of the technique, and the reasons for failure. Results: Responses were obtained from 104 of 324 (32.3%) identified emails. Responders were males (76%), seniors (59.7%), respiratory physicians (91.3%), working in a public/university hospital (78.8%), and mostly beginners (65.3%) from 41 countries. Following the course, 58.6% of responders either created or modified a MT program in their workplace. The reasons for not performing MT before the course were as follows: patients’ referral to a thoracic surgeon, not enough training, lack of funding, department understaffed, and refusal by the hospital/department. Overall, these reasons were significantly decreased (p = 0.002) after the course. Conclusions: Real-life data of our survey suggest that more than half of the responders have implemented the technique or modified their practice according to the skills they got from the course.
Preliminary Investigation about the Expression of G Protein-Coupled Receptors in Platelets from Patients with Chronic Immune Thrombocytopenic Purpura
Authors: Xu Daming , Xie Long , Zhang Zewen , Wang Duanxu , Qiu Jinfeng , Yu Wenjun , Xu Chengwei , He Chunling , Xu Xianru , Yin Jun
Published: April 2021

Objective: The objective of this study was to determine the expression of G protein-coupled receptors (GPCRs) in platelets from adult patients with chronic immune thrombocytopenic purpura (ITP). Methods: Peripheral blood samples were collected from 40 patients with chronic ITP in the Second Affiliated Hospital of Shantou University Medical College, and 40 peripheral blood samples from healthy volunteers were collected; expressions of the adenosine diphosphate receptors (P2Y1 and P2Y12), alpha-2A adrenergic receptor (α2A-AR), and thromboxane A2 receptor (TP) in platelets were detected by flow cytometry. Gα protein, protease-activated receptor 1 (PAR1), and protease-activated receptor 4 (PAR4) were analyzed by Western blot and analyzed statistically. Results: Flow cytometry measurements of mean fluorescence intensities showed platelets from patients with chronic ITP, compared to healthy individuals, had significantly higher levels of P2Y1 (31.4 ± 2.2 vs. 7.8 ± 0.8), P2Y12 (29.6 ± 2.1 vs. 7.2 ± 1.3), α2A-AR (25.8 ± 2.9 vs. 9.8 ± 0.9), and TP (39.8 ± 3.1 vs. 4.7 ± 0.6) (all p < 0.01). Similarly, integrated optical density analysis of Western blots showed that platelets from patients with chronic ITP had significantly higher levels of Gα (1046.3 ± 159.96 vs. 254.49 ± 39.51), PAR1 (832.98 ± 98.81 vs. 203.92 ± 27.47), and PAR4 (1518.80 ± 272.45 vs. 431.27 ± 41.86) (all p < 0.01). Conclusion: Expression of GPCRs is increased in platelets from patients with chronic ITP, suggesting that platelets of chronic ITP may participate in the complicated biological process by means of GPCR-mediated signaling pathways.
Protective Effects of Morus nigra and Its Phytochemicals against Hepatotoxicity: A Review of Preclinical Studies
Authors: Ghorbani Ahmad , Hooshmand Sara
Published: April 2021

Background: Our liver has a variety of vital functions including removing poisons, storing energy, immunological roles, and secretory and excretory functions. It may face some kinds of diseases caused by viruses, hepatotoxic chemicals, drugs, alcohol, and inherited disorders. Oxidative stress and inflammation are in the core of mechanisms of liver damages induced by viruses or chemical agents. Summary: Morus nigra (M. nigra), generally known as black mulberry, exhibited wide-spectrum pharmacological effects including antidiabetic, antinociceptive, anticancer, and hepatoprotective activities. Different parts of this plant particularly the fruit and leaf have shown beneficial effects on hepatocytes in cell culture and animal models of liver damages induced by chemicals (e.g., CCl4), drugs (e.g., paracetamol), diet (e.g., high fat), diabetes, etc. The beneficial effects of M. nigra on the liver are attributed to the presence of considerable amounts of phenolic compounds such as anthocyanins, flavonols, and phenolic acids. The present review is aimed to focus on the hepatoprotective activities of M. nigra and its phytochemicals and the mechanisms responsible for these activities. Key Messages: The evidence reviewed in this study can help design clinical trials on M. nigra in patients with liver disorders and develop a hepatoprotective herbal medicine.
Aluminum Microcomb Electrodes on Silicon Wafer for Detecting Val66Met Polymorphism in Brain-Derived Neurotrophic Factor
Authors: Li Zhi , Cui Liangmin , Zhao Hongyao , Du Jinxin , Gopinath Subash C.B. , Lakshmipriya Thangavel , Xin Xuezhi
Published: April 2021

Objective: Brain-derived neurotrophic factor (BDNF) dysregulation is widely related with various psychiatric and neurological disorders, including schizophrenia, depression, Rett syndrome, and addiction, and the available evidence suggests that BDNF is also highly correlated with Parkinson’s and Alzheimer’s diseases. Methods: The BDNF target sequence was detected on a capture probe attached on aluminum microcomb electrodes on the silicon wafer surface. A capture-target-reporter sandwich-type assay was performed to enhance the detection of the BDNF target. Results: The limit of detection was noticed to be 100 aM. Input of a reporter sequence at concentrations >10 aM improved the detection of the target sequence by enhancing changes in the generated currents. Control experiments with noncomplementary and single- and triple-mismatches of target and reporter sequences did not elicit changes in current levels, indicating the selective detection of the BDNF gene sequence. Conclusion: The above detection strategy will be useful for the detection and quantification of BDNF, thereby aiding in the provision of suitable treatments for BDNF-related disorders.