bluuebunny/crossref_metadata_2025 huggingface dataset:
Molecular Mechanism of Xixin-Ganjiang Herb Pair Treating Chronic Obstructive Pulmonary Disease-Integrated Network Pharmacology and Molecular Docking
Authors: Huang Ping , Huang Tao , Li Deshun , Han Lintao , Zhou Zhenxiang , Huang Fang , Li Jingjing , Wu Jiajia , Ye Yan , Wang Qiong , Duan Bailu
Published: June 2021

Background. Chronic obstructive pulmonary disease (COPD) is characterized by high morbidity, disability, and mortality, which seriously threatens human life and health. Xixin and Ganjiang are classic herb pairs of Zhongjing Zhang, which are often used to treat COPD in China. However, the substance basis and mechanism of action of Xixin-Ganjiang herb pair (XGHP) in the treatment of COPD remain unclear. Methods. On the website of TCMSP and the DrugBank, effective compounds and targets of XGHP were found. COPD targets were obtained from GeneCards, DisGeNET, and GEO gene chips. Intersecting these databases resulted in a library of drug targets for COPD. Then, intersection targets were used for protein-protein interaction (PPI) and pathway enrichment analysis. Finally, the binding activity between compounds and core genes was evaluated by molecular docking to verify the expression level of PTGS2 and PPARG in rats. Results. Twelve effective compounds and 104 core genes were found in the intersection library, and kaempferol, sesamin, β-sitosterol, PTGS2, and PPARG were particularly prominent in the network analysis. A total of 113 pathways were obtained and enrichment of the TNF signaling pathway, IL-17 signaling pathway, and C-type lectin receptor signaling pathway was particularly obvious. Molecular docking indicated that kaempferol, sesamin, and β-sitosterol were closely related to PTGS2 and PPARG and were superior to aminophylline. Key compounds in XGHP could restrict the expression of PTGS2 in the lung tissues of COPD rats and promote the expression of PPARG. Conclusion. Inhibition of the expression of inflammatory factor PTGS2 and promotion of the expression of PPARG may be an effective target of XGHP in the treatment of COPD.
Investigation on Wearable Antenna under Different Bending Conditions for Wireless Body Area Network (WBAN) Applications
Authors: Adam Ismahayati , Kamarudin Muhammad Ramlee , Rambe Ali H. , Haris Norshakila , Rahim Hasliza A. , Wan Muhamad Wan Zuki A. , Ismail Arif M. , Jusoh Muzammil , Yasin Mohd Najib M.
Published: June 2021

This paper analysed the effects of bending on the performance of a textile antenna wherein the antenna under test was made of felt substrate for both industrial, scientific, and medical (ISM) band and WBAN applications at 2.45 GHz. Moreover, the conductive material was used for the patch, and the ground plane used a 0.17 mm Shieldit textile. Meanwhile, the antenna structure was in the form of rectangular, with a line patch in between elements to abate the mutual coupling effect. The measured operating frequency range of the antenna spanned from 2.33 GHz to 2.5 GHz with a gain of 4.7 dBi at 2.45 GHz. In this paper, the antenna robustness was examined by bending the structure on different radii and degrees along both X- and Y-axis. Next, the effects on return loss, bandwidth, isolation, and radiation characteristics were analysed. This paper also discovered that the antenna’s performance remained acceptable as it was deformed, and the measured results agreed well with the simulation.
Investigation of Radiation-Induced Toxicity in Head and Neck Cancer Patients through Radiomics and Machine Learning: A Systematic Review
Authors: Carbonara Roberta , Bonomo Pierluigi , Di Rito Alessia , Didonna Vittorio , Gregucci Fabiana , Ciliberti Maria Paola , Surgo Alessia , Bonaparte Ilaria , Fiorentino Alba , Sardaro Angela
Published: June 2021

Background. Radiation-induced toxicity represents a crucial concern in oncological treatments of patients affected by head and neck neoplasms, due to its impact on survivors’ quality of life. Published reports suggested the potential of radiomics combined with machine learning methods in the prediction and assessment of radiation-induced toxicities, supporting a tailored radiation treatment management. In this paper, we present an update of the current knowledge concerning these modern approaches. Materials and Methods. A systematic review according to PICO-PRISMA methodology was conducted in MEDLINE/PubMed and EMBASE databases until June 2019. Studies assessing the use of radiomics combined with machine learning in predicting radiation-induced toxicity in head and neck cancer patients were specifically included. Four authors (two independently and two in concordance) assessed the methodological quality of the included studies using the Radiomic Quality Score (RQS). The overall score for each analyzed study was obtained by the sum of the single RQS items; the average and standard deviation values of the authors’ RQS were calculated and reported. Results. Eight included papers, presenting data on parotid glands, cochlea, masticatory muscles, and white brain matter, were specifically analyzed in this review. Only one study had an average RQS was ≤ 30% (50%), while 3 studies obtained a RQS almost ≤ 25%. Potential variability in the interpretations of specific RQS items could have influenced the inter-rater agreement in specific cases. Conclusions. Published radiomic studies provide encouraging but still limited and preliminary data that require further validation to improve the decision-making processes in preventing and managing radiation-induced toxicities.
Prognostic Value of Atherosclerotic Extent in Diabetic Patients with Nonobstructive Coronary Artery Disease
Authors: Ding Yipu , Liu Zinuan , Dou Guanhua , Yang Xia , Wang Xi , Shan Dongkai , He Bai , Jing Jing , Chen Yundai , Yang Junjie
Published: June 2021

Background and Objective. Atherosclerotic extent was proved to be associated with adverse cardiac events. Risk scores derived by coronary computed tomography angiography (CCTA) could identify high-risk group among patients with nonobstructive coronary artery disease (CAD), but the ability is still uncertain in the presence of diabetes mellitus (DM). The purpose of this study was to investigate the prognostic value of the atherosclerotic extent shown by CCTA in diabetic patients with nonobstructive CAD. Methods and Results. 813 DM patients (mean age

58.9
±
9.9

years, 48.1% male) referred for CCTA due to suspected CAD in 2015-2017 were consecutively included. During a median follow-up of 31.77 months, 50 major adverse cardiovascular events (MACEs) (6.15%) were experienced, including 2 cardiovascular deaths, 14 nonfatal myocardial infarctions, 27 unstable anginas requiring hospitalization, and 7 strokes. Three groups were defined based on coronary stenosis combined with Leiden score as normal, nonobstructive

Leiden
<
5

, and nonobstructive

Leiden
≥
5

. Cox models were used to assess the prognosis of plaque burden within these groups. An incremental incidence of MACE rates was observed. After adjustment for age, gender, and presence of high-risk plaque, the group of

Leiden
≥
5

showed a higher risk than

Leiden
<
5

(HR: 1.88, 95% CI: 1.03-3.42,

p
=
0.039

). Similar results were observed when segment involvement score (SIS) was used for sensitivity analysis. Conclusion. Atherosclerotic extent was associated with the prognosis of DM patients with nonobstructive coronary artery disease, highlighting the importance of better risk stratification and management.
Advances of Antisense Oligonucleotide Technology in the Treatment of Hereditary Neurodegenerative Diseases
Authors: Lin Mengsi , Hu Xinyi , Chang Shiyi , Chang Yan , Bian Wenjun , Hu Ruikun , Wang Jing , Zhu Qingwen , Qiu Jiaying
Published: June 2021

Antisense nucleic acids are single-stranded oligonucleotides that have been specially chemically modified, which can bind to RNA expressed by target genes through base complementary pairing and affect protein synthesis at the level of posttranscriptional processing or protein translation. In recent years, the application of antisense nucleic acid technology in the treatment of neuromuscular diseases has made remarkable progress. In 2016, the US FDA approved two antisense nucleic acid drugs for the treatment of Duchenne muscular dystrophy (DMD) and spinal muscular atrophy (SMA), and the development to treat other neurodegenerative diseases has also entered the clinical stage. Therefore, ASO represents a treatment with great potential. The article will summarize ASO therapies in terms of mechanism of action, chemical modification, and administration methods and analyze their role in several common neurodegenerative diseases, such as SMA, DMD, and amyotrophic lateral sclerosis (ALS). This article systematically summarizes the great potential of antisense nucleic acid technology in the treatment of hereditary neurodegenerative diseases.
Reducing Entropy Overestimation in Soft Actor Critic Using Dual Policy Network
Authors: Ali Hamid , Majeed Hammad , Usman Imran , Almejalli Khaled A.
Published: June 2021

In reinforcement learning (RL), an agent learns an environment through hit and trail. This behavior allows the agent to learn in complex and difficult environments. In RL, the agent normally learns the given environment by exploring or exploiting. Most of the algorithms suffer from under exploration in the latter stage of the episodes. Recently, an off‐policy algorithm called soft actor critic (SAC) is proposed that overcomes this problem by maximizing entropy as it learns the environment. In it, the agent tries to maximize entropy along with the expected discounted rewards. In SAC, the agent tries to be as random as possible while moving towards the maximum reward. This randomness allows the agent to explore the environment and stops it from getting stuck into local optima. We believe that maximizing the entropy causes the overestimation of entropy term which results in slow policy learning. This is because of the drastic change in action distribution whenever agent revisits the similar states. To overcome this problem, we propose a dual policy optimization framework, in which two independent policies are trained. Both the policies try to maximize entropy by choosing actions against the minimum entropy to reduce the overestimation. The use of two policies result in better and faster convergence. We demonstrate our approach on different well known continuous control simulated environments. Results show that our proposed technique achieves better results against state of the art SAC algorithm and learns better policies.
Microfragmented Adipose Tissue Injection (MFAT) May Be a Solution to the Rationing of Total Knee Replacement: A Prospective, Gender-Bias Mitigated, Reproducible Analysis at Two Years
Authors: Heidari Nima , Borg Tiffanie-Marie , Olgiati Stefano , Slevin Mark , Danovi Alessandro , Fish Brady , Wilson Adrian , Noorani Ali
Published: June 2021

Background. Knee osteoarthritis (KOA) is a significant cause of disability in a globally ageing population. Knee replacement surgery has been shown to improve function and quality of life. Access to this intervention can be limited for a number of reasons including rationing of care, lack of healthcare provision in austere environments, and more recently, due to the cessation of elective orthopaedic care as a result of the COVID pandemic. Referral for knee replacement surgery is often guided by the patient’s Oxford Knee Score (OKS). Recent therapies including treatment with microfragmented adipose tissue (MFAT) have emerged as alternatives to relieve pain and improve function in such patients. Method. We identified all patients with KOA Kellgren-Lawrence grade 3 and 4 in our dataset of patients treated with a single injection of MFAT and applied published OKS thresholds for referral for TKR to separate them into 3 cohorts according to their functional impairment. 220 patients (95 females, 125 males) with KOA were given one MFAT injection. The function (OKS) and quality of life (EuroQol-5) prior to and 24 months after therapy were compared. Results. MFAT injection provided a statistically significant improvement in the quality of life (EQ-5D) at 24 months in patients with a baseline OKS of 39 or less (

p

value: <0.001) as well as those with OKS of 27 or less who are deemed suitable for a knee replacement (

p

value: <0.001). Conclusion. MFAT injection improves quality of life in patients with KOA who are deemed suitable for the knee replacement. MFAT is a low-morbidity alternative biological treatment and can delay the need for total knee replacement in suitable patients.
An Experimental Study of the Influence of Hand‐Arm Posture and Grip Force on the Mechanical Impedance of Hand‐Arm System
Authors: Zhang Wenjie , Wang Qichao , Xu Zheng , Xu Hongmei , Li Hang , Dong Jiajun , Ma Xinbo
Published: June 2021

In order to investigate the effects of hand‐arm posture, grip force, push force, and vibration excitation intensity on the mechanical impedance of human hand‐arm system, a test system with a self‐developed vibration handle has been prepared. Based on the testing system, the mechanical impedance of the hand‐arm system of seven Chinese adult males were tested and calculated under the random vibration excitation with the frequency of 10–1000 Hz. The results reveal that when the frequency is lower (<40 Hz), the hand‐arm system with an elbow angle of 180o produces a higher mechanical impedance; when the frequency ranges from 40 Hz to 100 Hz, the hand‐arm system with an elbow angle of 90o generates a higher mechanical impedance; while when the frequency is higher (>100 Hz), the hand‐arm posture seems to have no obvious effect on the mechanical impedance. Higher grip or push force would increase the frequency corresponding to the peak value of the mechanical impedance and often correspond to a higher mechanical impedance in a specific frequency range (30–200 Hz). When the frequency is lower (<140 Hz), vibration intensity has certain effects on the mechanical impedance of the hand‐arm system. In conclusion, vibration intensity does not directly affect the mechanical impedance, but an increase in grip or push force often causes an increase in mechanical impedance and a higher frequency that corresponds to the peak of mechanical impedance.
Primary Analysis and 4-Year Follow-Up of the Phase III NIBIT-M2 Trial in Melanoma Patients With Brain Metastases
Authors: Di Giacomo Anna Maria , Chiarion-Sileni Vanna , Del Vecchio Michele , Ferrucci Pier Francesco , Guida Michele , Quaglino Pietro , Guidoboni Massimo , Marchetti Paolo , Cutaia Ornella , Amato Giovanni , Covre Alessia , Camerini Roberto , Calabrò Luana , Valente Monica , Giannarelli Diana , Mandalà Mario , Maio Michele
Published: June 2021

Purpose:

Phase II trials have shown encouraging activity with ipilimumab plus fotemustine and ipilimumab plus nivolumab in melanoma brain metastases. We report the primary analysis and 4-year follow-up of the NIBIT-M2 study, the first phase III trial comparing these regimens with fotemustine in patients with melanoma with brain metastases.

Patients and Methods:

This phase III study recruited patients 18 years of age and older with BRAF wild-type or mutant melanoma, and active, untreated, asymptomatic brain metastases from nine centers, randomized (1:1:1) to fotemustine, ipilimumab plus fotemustine, or ipilimumab plus nivolumab. The primary endpoint was overall survival (OS).

Results:

From January, 2013 to September, 2018, 27, 26, and 27 patients received fotemustine, ipilimumab plus fotemustine, and ipilimumab plus nivolumab. Median OS was 8.5 months [95% confidence interval (CI), 4.8–12.2] in the fotemustine arm, 8.2 months (95% CI, 2.2–14.3) in the ipilimumab plus fotemustine arm (HR vs. fotemustine, 1.09; 95% CI, 0.59–1.99; P = 0.78), and 29.2 months (95% CI, 0–65.1) in the ipilimumab plus nivolumab arm (HR vs. fotemustine, 0.44; 95% CI, 0.22–0.87; P = 0.017). Four-year survival rate was significantly higher for ipilimumab plus nivolumab than fotemustine [(41.0%; 95% CI, 20.6–61.4) vs. 10.9% (95% CI, 0–24.4; P = 0.015)], and was 10.3% (95% CI, 0–22.6) for ipilimumab plus fotemustine. In the fotemustine, ipilimumab plus fotemustine, and ipilimumab plus nivolumab arms, respectively, 11 (48%), 18 (69%), and eight (30%) patients had treatment-related grade 3 or 4 adverse events, without treatment-related deaths.

Conclusions:

Compared with fotemustine, ipilimumab plus nivolumab significantly improved overall and long-term survival of patients with melanoma with asymptomatic brain metastases.
Abstract NT-093: THE N6-METHYLATION OF ADENOSINE (M6A) IN FZD10 MRNA CONTRIBUTES TO RESISTANCE TO PARP INHIBITOR
Authors: Fukumoto Takeshi , Zhu Hengrui , Karakashev Sergey , Nacarelli Timothy , Fatkhutdinov Nail , Wu Shuai , Kossenkov Andrew V. , Showe Louise , Jean Stephanie , Zhang Lin , Zhang Rugang
Published: June 2021

Chemical modifications of RNAs have emerged as a new layer of epigenetic gene regulation. N6-methyladenosine (m6A) is the most abundant chemical modification of messenger RNA. The m6A modification affects RNA fate and functions such as RNA stability. Despite the high initial clinical response rates to PARP inhibitors in BRCA1/2-mutated epithelial ovarian cancers, PARP inhibitor (PARPi) resistance remains a major challenge. The role of m6A modification in PARPi resistance has not previously been explored. Here we show that m6A modification of FZD10 mRNA contributes to PARPi resistance by upregulating the Wnt/β-catenin pathway in BRCA-mutated EOC cells. Global m6A profile reveals a significant increase in m6A modification in FZD10 mRNA. This correlates with an increase in FZD10 mRNA stability and an upregulation of the Wnt/β-catenin pathway in PARPi resistant cells. FZD10 knockdown or inhibition of the Wnt/β-catenin sensitizes the resistant cells to PAPRi. Mechanistically, downregulation of m6A demethylases FTO and ALKBH5 is sufficient to increase FZD10 mRNA m6A modification and reduce PARPi sensitivity. Moreover, PAPRi and Wnt/β-catenin inhibitor showed synergistic suppression of growth of PAPRi resistant cancer both in vitro and in vivo in a xenograft ovarian cancer mouse model. Our results show that m6A contributes to PAPRi resistance in BRCA-deficient EOC cells by upregulating the Wnt/β-catenin pathway through stabilizing FZD10. They also suggest that inhibition of Wnt/β-catenin pathway represents a potential strategy to overcome PARPi resistance.

Citation Format: Takeshi Fukumoto, Hengrui Zhu, Sergey Karakashev, Timothy Nacarelli, Nail Fatkhutdinov, Shuai Wu, Andrew V. Kossenkov, Louise Showe, Stephanie Jean, Lin Zhang, Rugang Zhang. THE N6-METHYLATION OF ADENOSINE (M6A) IN FZD10 MRNA CONTRIBUTES TO RESISTANCE TO PARP INHIBITOR [abstract]. In: Proceedings of the 12th Biennial Ovarian Cancer Research Symposium; Sep 13-15, 2018; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2019;25(22 Suppl):Abstract nr NT-093.
Abstract NT-096: FABCLAVINE, A SECONDARY METABOLITE FROM XENORHABDUS BUDAPESTENSIS AS THERAPY AGAINST OVARIAN CANCER
Authors: Kapur Arvinder K , Kajla Mayur , Paskewitz Susan , Mehta Pooja , Mehta Geeta , Patankar Manish S
Published: June 2021

Ovarian cancer is the fifth most common cause of cancer-related deaths in women in the United States. Most women with ovarian cancer present with advanced disease. Even though initial response to surgery and chemotherapy is generally positive, many patients relapse with chemoresistant disease. We are especially interested in finding natural products that have anti-neoplastic activity against relapsed disease. Here, we report fabclavine, a secondary metabolite from Xenorhabdus budapestensis as novel anti-cancer agent.

Fabclavine was isolated and purified from cell-free supernatants of Xbu cultures. In a two-step procedure, active compounds were first precipitated from the supernatant with acetone and subsequently purified via reverse phase chromatography. Proliferation assays conducted on several ovarian cancer cell lines such as OVCAR3, OVCA433, ID8, IOSE, show that fabclavine inhibits the cancer cell proliferation at nanomolar (250-400 nM) concentration but has no effect on IOSE at that concentration. The ovarian cancer cell line OVCA33 when tested in proliferation assay showed, they are sensitive to fabclavine at a ten-fold lower concentration than cisplatin. Besides proliferation assay in 2D culture, we also tested the effect of fabclavine in 3D culture of ovarian cancer cell lines. The results indicate, fabclavine was equally effective in 3D culture and killed 40-50% of the cells in spheroids. In addition, fabclavine was also tested against cancer stem cells isolated from ovarian cancer patient and grown in 3D culture and it inhibited the growth of stem cells by 45%. Treatment of ovarian cancer cell lines with Fabclavine indicated DNA damage and an increase in cleaved caspase 3 and Annexin V binding, confirming that this agent was killing the OVCAR3, ID8 and OVCA433 cancer cell lines via apoptosis. Further experiments are underway to determine the apoptosis pathway (intrinsic or extrinsic) activated by fabclavine. Here we are describing for the first time a novel agent which is water soluble, active at low concentration and conveniently purified from cell free supernatant of bacterial culture. These properties of fabclavine makes it an excellent molecule to develop as therapy against ovarian tumors.

Citation Format: Arvinder K Kapur, Mayur Kajla, Susan Paskewitz, Pooja Mehta, Geeta Mehta, Manish S Patankar. FABCLAVINE, A SECONDARY METABOLITE FROM XENORHABDUS BUDAPESTENSIS AS THERAPY AGAINST OVARIAN CANCER [abstract]. In: Proceedings of the 12th Biennial Ovarian Cancer Research Symposium; Sep 13-15, 2018; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2019;25(22 Suppl):Abstract nr NT-096.
Abstract NT-108: TARGETING BROMODOMAIN PROTEINS AS A THERAPEUTIC STRATEGY FOR CLEAR CELL CARCINOMA OF THE OVARY
Authors: Shigeta Shogo , Shaw Reid , Moser Russell , Gurley Kay , Lui Goldie , Grandori Carla , Kemp Christopher
Published: June 2021

Background and Objectives: Clear cell carcinoma of the ovary (OCCC) is more resistant to platinum-based chemotherapy than high-grade serous ovarian cancer (HGSOC). Also, the indication for PARP inhibitors seems to be limited in OCCC because of the lower frequency of homologous recombination deficiency. As genomic background significantly differs among subtypes, therapeutic strategies should be optimized in response to each subtype or unique gene alterations. This study was designed to find novel therapeutic targets for OCCC by utilizing siRNA screens and unique patient-derived ovarian cancer cells that phenocopy original tumors.

Materials and Methods: Novel patient-derived ovarian cancer cells were established and kindly provided by Dr. Tan Ince at University of Miami. Genomic alterations of these cells were determined by DNA sequencing. siRNA screens targeting 2,172 unique genes were performed with ARID1A-mutated OCCC cell line OCI C5x, and high-grade serous ovarian cancer cell line OCI P5x. Genes inhibition of which significantly suppressed cell proliferation were selected as hits for each subtype independently. The difference of cell viability between OCI C5x and P5x was also compared, and subtype-specific hits were additionally determined. Hits were applied to Gene Ontology (GO) enrichment analysis to comprehend the gene set enrichment. Several prioritized targets for OCCC were validated with additional RNAi experiments and small molecule inhibitors.

Results: 114 and 108 genes were selected as hits specific to OCI C5x and OCI P5x, respectively. GO enrichment analysis revealed OCCC is potentially more vulnerable to epigenetic interventions than HOGSC. The following inspection revealed bromodomain-containing proteins were enriched in the hits for OCI C5x. Among these, we highlighted bromodomain and extra-terminal domain (BET) family proteins BRD2 and BRD3. Subsequent RNAi experiments validated BRD2 and BRD3 are highly potent therapeutic targets for OCCC.

Conclusions: siRNA screening results suggest different approaches are required according to subtypes for the targeted therapy of ovarian cancer. We propose that epigenetic intervention such as BET protein inhibition as a potential therapeutic option for OCCC, 50-60% of which harbor ARID1A mutation.

Citation Format: Shogo Shigeta, Reid Shaw, Russell Moser, Kay Gurley, Goldie Lui, Carla Grandori, Christopher Kemp. TARGETING BROMODOMAIN PROTEINS AS A THERAPEUTIC STRATEGY FOR CLEAR CELL CARCINOMA OF THE OVARY [abstract]. In: Proceedings of the 12th Biennial Ovarian Cancer Research Symposium; Sep 13-15, 2018; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2019;25(22 Suppl):Abstract nr NT-108.
Abstract NT-119: TARGET DISCOVERY OF NATURAL PRODUCT INSPIRED PHYLLANTHUSMINS FOR TREATMENT OF HIGH GRADE SEROUS OVARIAN CANCER
Authors: Young Alexandria N. , Kurina Steven , Huntsman Andrew , Rathnayake Rathnayake A. , Korkmaz Melissa , Kinghorn A. Douglas , Aldrich Leslie , Cologna Stephanie , Fuchs James R. , Burdette Joanna E.
Published: June 2021

BACKGROUND: High grade serous ovarian cancer (HGSOC) is a lethal gynecological malignancy with a need for new therapeutic agents. Many of the most widely used chemotherapeutic drugs are either derived from or are semi-synthetic derivatives of natural products. We developed potent synthetic analogs (PHYs) of the phyllanthusmin class inspired by prior natural product isolated from Phyllanthus poilanei Beille.

MATERIALS & METHODS: HGSOC cell lines, OVCAR3 and OVCAR8, and non-tumorigenic controls, IOSE80 and FT33, were used in this study. Cytotoxicity assays included sulforhodamine B assay, and annexin X/PI staining and Western blotting for confirmation of apoptosis induction. A photo affinity labeling method was used to attach PHY analogs to solid phase support. Targets were isolated using a pulldown technique and mass spectrometry. CRISPR-Cas9 genome editing was used to knockout and confirm putative targets.

RESULTS: The most potent analog, PHY34, has nanomolar potency in HGSOC cell lines in vitro and displayed cytotoxic activity through late-stage autophagy inhibition and activation of apoptosis. PHY34 was readily bioavailable through intraperitoneal administration in vivo where it significantly reduced HGSOC tumor burden. Targets were identified using photo affinity labeling-aided protein pulldown and mass spectrometry, and confirmed by generating knockout cell lines of targets.

CONCLUSIONS: This class of compounds holds promise as a potential, novel chemotherapeutic approach and demonstrates the effectiveness of pleiotropically targeting autophagy and apoptosis as a viable strategy for combating high grade serous ovarian cancer.

Citation Format: Alexandria N. Young, Steven Kurina, Andrew Huntsman, Rathnayake A. Rathnayake, Melissa Korkmaz, A. Douglas Kinghorn, Leslie Aldrich, Stephanie Cologna, James R. Fuchs, Joanna E. Burdette. TARGET DISCOVERY OF NATURAL PRODUCT INSPIRED PHYLLANTHUSMINS FOR TREATMENT OF HIGH GRADE SEROUS OVARIAN CANCER [abstract]. In: Proceedings of the 12th Biennial Ovarian Cancer Research Symposium; Sep 13-15, 2018; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2019;25(22 Suppl):Abstract nr NT-119.
Abstract NT-120: KNOCKOUT OF MTF1 RESULTS IN THE INHIBITION OF EMT IN OVARIAN CANCER CELLS
Authors: Zhao Guannan , Yue Lawrence M Pfeffer Junming
Published: June 2021

Due to peritoneal metastasis and frequent recurrence, ovarian cancer has the highest mortality among gynecological cancers. Epithelial to mesenchymal transition (EMT) contributes to ovarian tumor metastasis. In this study, we report for the first time that metal regulatory transcription factor 1 (MTF1) was upregulated in ovarian cancer, and its high expression was associated with poor patient survival and disease relapse. Knockout of MTF1 using lentiviral CRISPR/Cas9 nickase vector-mediated gene editing inhibited EMT by upregulating epithelial cell markers E-cadherin and cytokeratin 7, and downregulating mesenchymal markers Snai2 and β-catenin in ovarian cancer SKOV3 and OVCAR3 cells. Loss of MTF1 reduced cell proliferation, migration, and invasion in both SKOV3 and OVCAR3 cells. Knockout of MTF1 upregulated the expression of the KLF4 transcription factor, and attenuated two cellular survival pathways, ERK1/2 and AKT. Our studies demonstrated that MTF1 plays an oncogenic role and contributes to ovarian tumor metastasis by promoting EMT. MTF1 may be a novel biomarker for early diagnosis as well as a drug target for clinical therapy.

Citation Format: Guannan Zhao, Lawrence M Pfeffer Junming Yue. KNOCKOUT OF MTF1 RESULTS IN THE INHIBITION OF EMT IN OVARIAN CANCER CELLS [abstract]. In: Proceedings of the 12th Biennial Ovarian Cancer Research Symposium; Sep 13-15, 2018; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2019;25(22 Suppl):Abstract nr NT-120.
Abstract TMIM-078: USING A DYNAMIC, LOW-SHEAR ENVIRONMENT TO MIMIC PHYSIOLOGICAL FLUID SHEAR STRESS IN OVARIAN CANCER IN VITRO MODELING
Authors: Masiello Timothy J. , Hemachandra L.P. Madhubhani , Castracane James
Published: June 2021

Ovarian cancer is the most lethal gynecological cancer with a five-year survival rate as low as 30%. Early diagnosis could offer high chances of survival as most ovarian cancer cases are diagnosed only after metastasis has taken place. Some of the challenges in diagnosis and treatment are due to ovarian cancer's preference for the transcoelomic metastatic pathway over the more well-known hematogenic pathway, where spheroids suspended in the peritoneal cavity invade through the mesothelial layer that lines it. Spheroids for in vitro experiments have been generated for decades using common methods such as hanging drops and liquid overlay, but these models typically do not account for the fluid shear stress present in vivo, especially following the formation of ascites. One method to mimic this in vitro is to use dynamic cell culture to generate shear stress. Fluid movement parameters (rotation, shaking, etc.) can be tuned to achieve physiologically-relevant shear stress while facilitating the formation of spheroids. Dynamic culture improves the roundness and size consistency of spheroids, facilitating functional assays such as spheroid migration and mesothelial clearance. The addition of shear stress is expected to increase the metastatic behavior of spheroids as it better recreates the in vivo environment. This model also provides a platform for incorporating other in vivo elements such as co-culture, use of ascites samples, etc. This work is ultimately aimed at improving our fundamental understanding of peritoneal metastasis with new insights for determining optimal therapeutic approaches.

Citation Format: Timothy J. Masiello, L.P. Madhubhani Hemachandra, and James Castracane. USING A DYNAMIC, LOW-SHEAR ENVIRONMENT TO MIMIC PHYSIOLOGICAL FLUID SHEAR STRESS IN OVARIAN CANCER IN VITRO MODELING [abstract]. In: Proceedings of the 12th Biennial Ovarian Cancer Research Symposium; Sep 13-15, 2018; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2019;25(22 Suppl):Abstract nr TMIM-078.
Comprehensive Genomic and Transcriptomic Analysis for Guiding Therapeutic Decisions in Patients with Rare Cancers
Authors: Horak Peter , Heining Christoph , Kreutzfeldt Simon , Hutter Barbara , Mock Andreas , Hüllein Jennifer , Fröhlich Martina , Uhrig Sebastian , Jahn Arne , Rump Andreas , Gieldon Laura , Möhrmann Lino , Hanf Dorothea , Teleanu Veronica , Heilig Christoph E. , Lipka Daniel B. , Allgäuer Michael , Ruhnke Leo , Laßmann Andreas , Endris Volker , Neumann Olaf , Penzel Roland , Beck Katja , Richter Daniela , Winter Ulrike , Wolf Stephan , Pfütze Katrin , Geörg Christina , Meißburger Bettina , Buchhalter Ivo , Augustin Marinela , Aulitzky Walter E. , Hohenberger Peter , Kroiss Matthias , Schirmacher Peter , Schlenk Richard F. , Keilholz Ulrich , Klauschen Frederick , Folprecht Gunnar , Bauer Sebastian , Siveke Jens Thomas , Brandts Christian H. , Kindler Thomas , Boerries Melanie , Illert Anna L. , von Bubnoff Nikolas , Jost Philipp J. , Spiekermann Karsten , Bitzer Michael , Schulze-Osthoff Klaus , von Kalle Christof , Klink Barbara , Brors Benedikt , Stenzinger Albrecht , Schröck Evelin , Hübschmann Daniel , Weichert Wilko , Glimm Hanno , Fröhling Stefan
Published: June 2021

The clinical relevance of comprehensive molecular analysis in rare cancers is not established. We analyzed the molecular profiles and clinical outcomes of 1,310 patients (rare cancers, 75.5%) enrolled in a prospective observational study by the German Cancer Consortium that applies whole-genome/exome and RNA sequencing to inform the care of adults with incurable cancers. On the basis of 472 single and six composite biomarkers, a cross-institutional molecular tumor board provided evidence-based management recommendations, including diagnostic reevaluation, genetic counseling, and experimental treatment, in 88% of cases. Recommended therapies were administered in 362 of 1,138 patients (31.8%) and resulted in significantly improved overall response and disease control rates (23.9% and 55.3%) compared with previous therapies, translating into a progression-free survival ratio >1.3 in 35.7% of patients. These data demonstrate the benefit of molecular stratification in rare cancers and represent a resource that may promote clinical trial access and drug approvals in this underserved patient population.

Significance:

Rare cancers are difficult to treat; in particular, molecular pathogenesis–oriented medical therapies are often lacking. This study shows that whole-genome/exome and RNA sequencing enables molecularly informed treatments that lead to clinical benefit in a substantial proportion of patients with advanced rare cancers and paves the way for future clinical trials.

See related commentary by Eggermont et al., p. 2677.

This article is highlighted in the In This Issue feature, p. 2659
Abstract A19: Metronomic chemotherapy enhances immunotherapy by preventing stroma-induced immunosuppression
Authors: Tsai Kelvin K. , Chan Tze-sian
Published: June 2021

Chemotherapy (C/T) drugs are usually administered to cancer patients every few weeks at a high, “maximum tolerated” dose (MTD). Though this approach kills the majority of tumor cells, it often spares a small number of stem-like cancer cells (CSCs) that subsequently give rise to new tumors. Moreover, these recurring tumors are often more aggressive and able to metastasize to other tissues, in part because high doses of C/T drugs also affect cells in the stromal tissue that surrounds tumors, including carcinoma-associated fibroblasts (CAFs) and immune cells. Many human solid tumors are characterized by a pronounced stromal reaction characterized by a desmoplastic response produced by CAFs as well as an extensive infiltration by immunosuppressive cell populations such as tumor-associated macrophages (TAMs) and myeloid-derived suppressor cells (MDSCs). Importantly, both the tumor epithelium and adjacent stroma respond to systemic C/T and a therapy-modified stroma can deleteriously influence treatment efficacy. Recently, we have demonstrated in assorted mouse tumor models that traditional C/T using the MTD regimens of C/T agents such as cyclophosphamide, doxorubicin, gemcitabine, and paclitaxel could paradoxically render CAFs oncogenic through the stromal-epithelial ELR+-chemokine/CXCR-2 signaling axis, which expands CSCs, enhances tumor neovascularization, and induces immunosuppressive M2 subtype TAM infiltration, together leading to tumor recurrence and treatment refractoriness (Chan et al., J. Exp. Med 2016). By contrast, the same total accumulated dose of the C/T agents administered as a low-dose metronomic (LDM) regimen could largely prevent the therapy-induced stromal alterations and CSC expansion and thereby substantially enhance the treatment response. We subsequently showed that LDM C/T not only obviated the MTD-C/T-elicited CSC expansion but also attenuated the influx of the highly immunosuppressive granulocytic MDSCs, thereby potentiating immunotherapy such as anti-PD-1. We thus anticipate that, while pharmacologic inhibition of CXCR-2 has enhanced the antitumor efficacy of immune checkpoint blockade, LDM C/T may provide a clinically applicable alternative of enhancing immune surveillance in stroma-rich and T-cell inflamed tumors. Collectively, our studies provide compelling evidence and novel mechanistic insights into the unique benefit of LDM C/T on the tumor stroma. Given that MTD C/T exerts favorable influences on tumor stemness and immunity, it may better synergize with emerging CSC-targeted or immunotherapeutics than traditional MTD C/T to further enhance the therapeutic outcome of human solid tumors.

Citation Format: Kelvin K. Tsai, Tze-sian Chan. Metronomic chemotherapy enhances immunotherapy by preventing stroma-induced immunosuppression [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2018 Nov 27-30; Miami Beach, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(4 Suppl):Abstract nr A19.
Abstract A24: Antigen-targeted soluble bispecific T-cell receptor (ImmTACTM) molecules in immunotherapy
Authors: Vuidepot Annelise
Published: June 2021

Cancer immunotherapies function to harness the body’s own immune system to eradicate cancer cells. Critical to this process is the T cell, capable of directing potent and antigen-specific immune responses. T-cell receptor (TCR) antigens are short intracellularly processed peptides presented on the cell surface. As the majority of the proteome is processed and presented on the surface of cells and hence can be targeted by a TCR, TCR-based therapies offer distinct advantages over antibody-based therapies that are limited to targeting only secreted or cell surface proteins. In this study, we describe our approach to address the limitations and utilize the natural advantages of the human TCR to target a diverse range of tumor indications. Immune mobilizing monoclonal TCRs Against Cancer (ImmTAC) molecules are a novel class of bispecific biologic formed from an affinity-engineered, soluble, targeted TCR fused to a T cell redirecting anti-CD3 scFv. We describe our process to engineer high-affinity ImmTAC molecules towards novel cancer antigens that have been identified and validated in house. Additionally, we discuss the challenges and opportunities in the preclinical testing of an exquisitely human-specific biologic that led to the development of an entirely in vitro preclinical package that is proven to support the clinical development of ImmTAC molecules to clinical development. In summary, we present a novel approach to cancer immunotherapy, detailing the engineering, optimization, preclinical testing and clinical development of ImmTAC molecules, a new bispecific biologic.

Note: This abstract was not presented at the conference.

Citation Format: Annelise Vuidepot. Antigen-targeted soluble bispecific T-cell receptor (ImmTACTM) molecules in immunotherapy [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2018 Nov 27-30; Miami Beach, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(4 Suppl):Abstract nr A24.
Abstract A31: Comparing IL-12 and IL-18 secreting CAR T-cell efficacy
Authors: Bebernitz Christina E. , Brentjens Renier J.
Published: June 2021

MUC16, a subunit of CA125 that is overexpressed on epithelial ovarian cancers, can be targeted by T cells via the incorporation of the 4h11m28m chimeric antigen receptor (CAR) into the T-cell surface. However, the microenvironment of ovarian cancers consists of regulatory T cells, myeloid-derived suppressor cells, and tumor-associated macrophages that act to immunosuppress CAR T-cell function, thus abrogating CAR T-cell efficacy. To overcome this immunosuppression, our lab has developed IL-12 and IL-18 secreting CAR T cells, which have shown enhanced efficacy in a syngeneic ovarian cancer model when compared to second-generation CAR T cells. While IL-18 secreting CAR T cells have proven to be superior to IL-12 secreting CAR T cells in various leukemia and lymphoma mouse models, this observation does not translate to the ovarian cancer model. Mechanistic studies into the effects of IL-18 and IL-12 in a syngeneic ovarian cancer model are currently under way in order to better understand what each cytokine is doing to the tumor microenvironment as well as to the CAR T cell. With better understanding of the effects of IL-18 and IL-12, we can contribute to the immunobiology of these cytokines as well as harness their efficacy to apply to other tumor models.

Citation Format: Christina E. Bebernitz, Renier J. Brentjens. Comparing IL-12 and IL-18 secreting CAR T-cell efficacy [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2018 Nov 27-30; Miami Beach, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(4 Suppl):Abstract nr A31.
Abstract A61: The positive effect of SKD plus IOT and HBO2T in the treatment of cancer
Authors: Niknamian Sorush
Published: June 2021

Background: The aim of this research is to figure out the effectiveness of the Sorush Cancer Treatment Protocol (SCTP), which is based on the Evolutionary Metabolic Hypothesis of Cancer (EMHC) and introducing the Specific Ketogenic Diet (SKD) plus Intravenous Ozone Therapy (IOT) in phase (1) on 54 cancer patients, and combination of hyperbaric oxygen therapy with vitamin/mineral and herbal supplementation beside the SKD and IOT in phase (2) of this research on the remaining 31 cancer patients.

Introduction: Cancer based on the introduction of EMHC by Dr. Somayeh Zaminpira and Dr. Sorush Niknamian in 2017 is an evolutionary metabolic disease and through the incline of the reactive oxygen species (ROS) and the Butterfly Effect in normal eukaryotic cells, the mitochondria become damaged or shut down. Cancer cells are primitive eukaryotic cells that have existed since around 1.5 billion years ago before the entrance of mitochondrion as endosymbiont.

Materials and Methods: Based on the researches from 1928-2016 and the experimentation of cancer treatments and protocols on cancer patients, we have reached a treatment and decided to test it on 54 voluntary cancer patients in the first stage of their disease. In this treatment we used a 5-day water fasting state, the Specific Ketogenic Diet (SKD) designed by ourselves and Intravenous Ozone Therapy (IOT) in the duration of 90 days (phase 1) and another 90 days (phase 2) with the entrance of hyperbaric oxygen therapy (HBO2T) and several supplements that have been effective in previous studies on cancer patients. We have used the measurement of saliva PH, the MRI device and statistical methods to test the shrinkage of the tumors.

Results: After phase (1) of this research on 54 patients, the average percentage decrease in the tumors was 58%, and after phase (2) on 31 remaining cancer patients the average percentage decrease in the tumors was 98.8%. The average saliva PH in the fasting state of the cancer patients improved from acidic to alkaline as well.

Conclusion: We have reached an effective cancer treatment based on SCTP by the usage of SKD, IOT, HBO2T and several supplements. There was an obvious improvement of cancer tumor decrease, lifestyle, and saliva PH, and we did not observe any side effects or cachexia in any of the patients.

Citation Format: Sorush Niknamian. The positive effect of SKD plus IOT and HBO2T in the treatment of cancer [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2018 Nov 27-30; Miami Beach, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(4 Suppl):Abstract nr A61.
Abstract A62: Directly visualizing interactions between lung microbiota, tissue-resident immune cells and tumor cells in a genetically engineered lung adenocarcinoma mouse model
Authors: Zhao Chen , Li Weizhe , Jin Chengcheng , Germain Ronald N.
Published: June 2021

Lung adenocarcinoma is closely associated with chronic inflammation, but the underlying mechanism has not been fully elucidated yet. As a mucosal organ, lung was thought to be sterile in the past due to the limitation of bacteria culture techniques, but recent 16s RNA sequencing analysis has proved that lung is colonized by a diverse bacterial community. Recent studies suggest that the dysbiosis of lung microbiota plays a critical role in lung cancer tumorigenesis. However, there is no direct evidence addressing the complex interactions between local microbiota, immune cells and tumor cells due to the limitation of traditional imaging methods. Here we developed an advanced imaging technique combining RNA fluorescent in situ hybridization, immunostaining and Clearing-enhanced 3D (Ce3D), and applied it in a genetically engineered mouse model harboring the most common mutations (activation of Kras and p53 loss of function) in patients with lung adenocarcinoma. It revealed a strong spatial-temporal correlation between lung microbiota and tissue-resident immune cells during lung adenocarcinoma development. This research was supported by the Intramural Research Programs of NIAID and NHLBI, NIH.

Citation Format: Chen Zhao, Weizhe Li, Chengcheng Jin, Ronald N. Germain. Directly visualizing interactions between lung microbiota, tissue-resident immune cells and tumor cells in a genetically engineered lung adenocarcinoma mouse model [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2018 Nov 27-30; Miami Beach, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(4 Suppl):Abstract nr A62.
Abstract A64: Commensal dysbiosis alters the tumor microenvironment in breast cancer and diminishes efficacy of PD-L1 blockade
Authors: Rosean Claire Buchta , Bostic Raegan , Feng Tzu Yu , Rutkowski Melanie R.
Published: June 2021

Hormone receptor-positive (HR+) breast cancer is largely nonresponsive to checkpoint blockade therapy. Alterations in the composition of commensal microbes influence the systemic immune environment and can diminish responses to checkpoint blockade. Reduced diversity of commensal microbes, known as dysbiosis, has been demonstrated in women with advanced breast cancer and is associated with increases of systemic immunosuppressive mediators that are detrimental in the context of breast cancer, including prostaglandin E2 (PGE2). This bioactive lipid promotes the differentiation of myeloid cells that can exert immunosuppressive effects in the tumor microenvironment. In a novel, clinically relevant model of HR+ breast cancer, PD-L1 blockade was effective when combined with meloxicam, a drug that targets PGE2 synthesis through inhibition of COX-2. However, this combination therapy was ineffective in mice that were previously treated with antibiotics to induce dysbiosis, despite elevated levels of PGE2 in these animals. Commensal dysbiosis altered the cellular composition within the tumor microenvironment, promoting the recruitment of suppressive myeloid populations and driving significant and irreversible T-cell dysfunction. Importantly, COX-1 also regulates PGE2 synthesis, and mammary glands from dysbiotic mice showed significantly enhanced COX-1 activity, suggesting a previously unknown mechanism by which dysbiosis can affect breast tumor progression and resistance to therapy. Targeting COX-1-mediated PGE2 synthesis resulted in decreased tumor burden only in dysbiotic mice, suggesting that dysbiosis-induced COX-1-mediated tissue inflammation is sufficient to drive immune suppression in advanced breast cancer. This work provides important insights on the premalignant environment in the breast, enhances our understanding of the mechanisms driving failure of PD-1/PD-L1 blockade, and identifies novel targets that may provide clinical benefit for women with breast cancer.

Citation Format: Claire Buchta Rosean, Raegan Bostic, Tzu Yu Feng, Melanie R. Rutkowski. Commensal dysbiosis alters the tumor microenvironment in breast cancer and diminishes efficacy of PD-L1 blockade [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2018 Nov 27-30; Miami Beach, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(4 Suppl):Abstract nr A64.