bluuebunny/crossref_metadata_2025 huggingface dataset:
Successful Management of Infectious Crystalline Keratopathy with Intrastromal Antibiotic Injections
Authors: Martinez-Velazquez Luis , Ma Kevin K. , Patel Neal S. , Luo Zhonghui Katie
Published: December 2022

Purpose. To report the successful treatment of a case of presumed infectious crystalline keratopathy with repeated intrastromal antibiotic injections in a cornea graft in the setting of severe ocular graft-vs.-host-disease (GVHD). Observations. A 62-year-old man with a history of ocular GVHD and tectonic penetrating keratoplasty (PK) for corneal melt from herpes zoster keratopathy developed presumed infectious crystalline keratopathy (ICK) in the corneal graft. Given the patient’s complicated ocular history, chronic immunosuppression and new cardiac comorbidities, a therapeutic PK would most likely fail. Efforts were then directed to rescue the graft with minimally invasive approaches. Two separate intrastromal injections of cefuroxime and moxifloxacin successfully treated his ICK. Conclusions and Importance. This case supports a role for repeated intrastromal antibiotic injections in patients with ICK refractory to topical antibiotic therapy, which might eliminate the need for therapeutic PK and preserve vision.
Diagnostic Value of Metagenomic Next-Generation Sequencing for Pneumonia in Immunocompromised Patients
Authors: Li Jun , Zhou Chao-E. , Wei Shan-Chen , Wang Li-Na , Shi Ming-Wei , Sun Chun-Ping , Lin Lian-Jun , Liu Xin-Min
Published: December 2022

Introduction. The diagnosis of pulmonary infection and the identification of pathogens are still clinical challenges in immunocompromised patients. Metagenomic next-generation sequencing (mNGS) has emerged as a promising infection diagnostic technique. However, its diagnostic value in immunocompromised patients needs further exploration. Purposes. This study was to evaluate the diagnostic value of mNGS compared with comprehensive conventional pathogen tests (CTs) in the etiology of pneumonia in immunocompromised patients and immunocompetent patients. Methods. We retrospectively reviewed 53 patients who were diagnosed with pneumonia from May 2019 to June 2021. There were 32 immunocompromised patients and 21 immunocompetent patients with pneumonia who received both mNGS and CTs. The diagnostic performance was compared between mNGS and CTs in immunocompromised patients, using the composite diagnosis as the reference standard. And, the diagnostic value of mNGS for mixed infections was further analyzed. Results. Compared to immunocompetent patients, the most commonly pathogens, followed by Cytomegalovirus, Pneumocystis jirovecii and Klebsiella pneumoniae in immunocompromised patients. Furthermore, more mixed infections were diagnosed, and bacterial-fungal-virus coinfection was the most frequent combination (43.8%). mNGS can detect more types of pathogenic microorganisms than CTs in both groups (78.1% vs. 62.5%,

P
=
0.016

and 57.1% vs. 42.9%,

P
=
0.048

). The overall diagnostic positive rate of mNGS for pathogens was higher in immunocompromised patients (

P
=
0.002

). In immunocompromised patients, a comparable diagnostic accuracy of mNGS and CTs was found for bacterial, fungal, and viral infections and coinfection. mNGS had a much higher sensitivity for bacterial infections (92.9% vs. 50%,

P
<
0.001

) and coinfections (68.8% vs. 48.3%,

P
<
0.05

), and it had no significant advantage in the detection of fungal infections, mainly due to the high sensitivity for Pneumocystis jirovecii in both groups. Conclusion. mNGS is more valuable in immunocompromised patients and exhibits apparent advantages in detecting bacterial and mixed infections. It may be an alternative or complementary diagnostic method for the diagnosis of complicated infections in immunocompromised patients.
Serum LAPTM4B as a Potential Diagnostic and Prognostic Biomarker for Breast Cancer
Authors: Wang Lu , Wang Yating , Zhang Qingyun
Published: December 2022

Background. Lysosome‐associated protein transmembrane‐4 beta (LAPTM4B) is an integral membrane protein overexpressed in various cancers and may function as a prognostic tumor marker. The present study is aimed at understanding the clinical significance of serum LAPTM4B in breast cancer (BC). Methods. Serum LAPTM4B level was evaluated in 426 BC patients, 40 benign breast disease, and 80 healthy controls by ELISA. We used the receiver operator characteristic (ROC) curve to assess the diagnostic significance. 46 BC patients were recruited to monitor the dynamic change of serum LAPTM4B during adjuvant therapy (AT). In addition, sera from a subset of 330 patients undergoing AT, including anti‐HER2 treatment, were collected to evaluate the association between LAPTM4B levels and AT efficacy. Descriptive and explorative statistical analyses were used to assess LAPTM4 B’s potential as a diagnostic and prognostic marker in BC. Results. Serum LAPTM4B level was significantly increased in BC patients than benign group and controls. It could well discriminate BC from healthy controls with diagnostic accuracy with an AUC of 0.912, a sensitivity of 85.9%, and a specificity of 83.8%. Compared with pre‐AT, serum LAPTM4B concentration remarkably decreased after AT. In addition, patients in the invalid response group (PD + SD) showed higher LAPTM4B levels than the valid response group (PR + CR). Conclusion. Our results proposed that serum LAPTM4B had a high diagnostic and prognostic impact as a circulating biomarker in BC.
Everolimus for Estrogen Receptor–negative Breast Cancer Prevention: A Journey Begun?
Authors: Nye Lauren , Khan Seema A.
Published: December 2022

Tamoxifen, the prototypic medication for breast cancer prevention, was approved for this purpose by the FDA in 1998. Other drugs have been proven to be effective in the ensuing decades. But the two major limitations of these have become clear over time: a lack of protection against hormone receptor–negative breast cancer, and a profile of safety and tolerability that is unacceptable to the majority of women at increased breast cancer risk. Recent preclinical data on targeting of the key oncogenic pathway of PI3K/AKT/mTOR signaling with drugs such as rapamycin and everolimus are provocative. Their efficacy signal should be pursued with further research, but their safety and tolerability profiles remain a concern.

See related article by Mazumdar et al., p. 791
Abstract B003: Reprogramming of fibrotic macrophages to control DCIS progression
Authors: Hayward Mary-Kate , Timblin Greg A. , Northey Jason J. , Stashko Connor , Ironside Alastair J. , Hwang E. Shelley , Weaver Valerie M.
Published: December 2022

Women with aggressive DCIS are at high risk of progression to invasive breast cancer. Invasive disease is accompanied by extensive collagen remodeling and crosslinking that contributes to tissue fibrosis and stiffness. Fibrotic breast tumors are associated with an immune desert or immune excluded tumor microenvironment (TME) that functions to promote tumor progression. In particular, tumor-associated macrophages (TAMs) promote tumor invasion and metastasis and, drive immune suppression allowing tumors to evade intrinsic anti-tumor immune responses. Here, we show that collagen remodeling and crosslinking and tissue stiffness associate with DCIS progression and recurrence to invasion in patient tissues. The fibrotic and stiff TME promotes DCIS progression in a mouse model in part by inducing a suppressive TAM phenotype characterized by epigenetic and metabolic reprogramming that accompanies an impaired pro-inflammatory gene signature, as seen in tolerant macrophages. TAMs can be polarized by toll-like receptor (TLR) agonists to eliminate tumor cells by producing pro-inflammatory cytokines or by stimulating cytotoxic T cells, though these are poor cancer therapies alone. Thus, we hypothesized to improve the TLR agonists anti-tumor effects, we can reprogram the metabolic pathways associated with macrophage tolerance, and these together may control DCIS progression in fibrotic and stiff TMEs. We show that fibrotic TAMs can be reprogrammed with monophosphoryl lipid A (MPLA) and coenzyme A (CoA) to kill tumor cells. MPLA + CoA injected into the mammary fat pad reduces high risk features of DCIS in this breast cancer mouse model. We show that CoA supplementation alters macrophage metabolism by supporting acetyl-CoA production needed for histone acetylation and pro-inflammatory gene transcription. MPLA + CoA reprograms CD206+ TAMs to inducible NO synthase (iNOS)+ macrophages and activates cytotoxic T cells through macrophage-secreted cytokines. This treatment approach for engaging an anti-tumor immune response represents an alternative treatment to that of current invasive surgery options for women with high-risk DCIS.

Citation Format: Mary-Kate Hayward, Greg A. Timblin, Jason J. Northey, Connor Stashko, Alastair J. Ironside, E. Shelley Hwang, Valerie M. Weaver. Reprogramming of fibrotic macrophages to control DCIS progression [abstract]. In: Proceedings of the AACR Special Conference on Rethinking DCIS: An Opportunity for Prevention?; 2022 Sep 8-11; Philadelphia, PA. Philadelphia (PA): AACR; Can Prev Res 2022;15(12 Suppl_1): Abstract nr B003.
Abstract B009: A SOX9-B7x axis safeguards dedifferentiated tumor cells from immunosurveillance to enable DCIS progression
Authors: Liu Yu , John Peter , Christin John R , Oktay Maja , Zang XingXing , Guo Wenjun
Published: December 2022

Accumulating evidence has demonstrated a central role of dedifferentiation in carcinogenesis. In basal-like breast cancer mouse models, upregulation of SOX9 drives dedifferentiation of luminal progenitors to cells with luminal-basal hybrid features resembling embryonic multipotent mammary stem cells. Additionally, genetic inactivation of SOX9 inhibits dedifferentiation and blocks the progression of in situ tumors to malignant carcinoma. Whether the host immune system can recognize these dedifferentiated cells and how they eventually evade immunosurveillance are unclear. Here, we showed that SOX9 was necessary for limiting infiltrating T lymphocytes to protect dedifferentiated cells from immune surveillance in premalignant lesions of basal-like breast cancer. SOX9 deletion in spontaneous BLBC tumor models caused massive T lymphocyte infiltration and elimination of dedifferentiated cells. Antibody-mediated T cell depletion rescued the accumulation of dedifferentiated tumor cells in SOX9-deficient DCIS and its progression to invasive tumors. Mechanistically, through both stimulation of the STAT3 pathway and direct transcriptional activation, SOX9 upregulated the expression of the immune checkpoint B7x, which mediated the action of SOX9 in immunosuppression. In advanced tumors, B7x blockade reactivated antitumor immunity and effectively inhibited tumor growth. Furthermore, B7x depletion overcame the resistance to anti-PD-L1 immunotherapy. Collectively, these findings uncovered a dedifferentiation-associated immune evasion mechanism and demonstrated the therapeutic potential of targeting the SOX9-B7x pathway to eradicate dedifferentiated tumor cells for intercepting breast cancer progression.

Citation Format: Yu Liu, Peter John, John R Christin, Maja Oktay, XingXing Zang, Wenjun Guo. A SOX9-B7x axis safeguards dedifferentiated tumor cells from immunosurveillance to enable DCIS progression [abstract]. In: Proceedings of the AACR Special Conference on Rethinking DCIS: An Opportunity for Prevention?; 2022 Sep 8-11; Philadelphia, PA. Philadelphia (PA): AACR; Can Prev Res 2022;15(12 Suppl_1): Abstract nr B009.
Abstract B018: Physical tumor microenvironment biomarkers to predict DCIS progression, upstaging, and early metastasis
Authors: Damaghi Mehdi , Raza Zainab , Farman Rana Adil , Bai JiDong , Saltz Joel , Niell Bethany L.
Published: December 2022

Ductal carcinoma in situ (DCIS) is defined as a neoplastic proliferation of epithelial cells confined to the mammary ductal-lobular system. Although metastasis can occur at the very early stages of DCIS, the majority of metastatic breast cancers are believed to arise through a years-long process of somatic evolution within the physical constraints of milk ducts, characterized by the accumulation of heritable genetic and epigenetic changes. While these genetic alterations during breast carcinogenesis have been extensively studied without any outcome that can be used as a clinical biomarker, less attention has been paid to the microenvironmental conditions that select for these genotypes. We have demonstrated that evolving pre-cancerous hyperplastic breast cells are selected under physical tumor microenvironment conditions such as hypoxia, acidosis, and starvation. We have also shown that these harsh conditions select cells with more aggressive phenotypes. We have developed several markers for the adapted cells under these harsh conditions such as LAMP2b for acidosis or CA9 for hypoxia. We, therefore, hypothesize that the expression of physical microenvironment adaptation proteins (PMA) can differentiate indolent from aggressive DCIS and that this can be used for decision support. DCIS can be detected by screening mammography followed by a percutaneous Biopsy for pathologic confirmation, which may be followed by surgery +/- adjuvant therapy.

Citation Format: Mehdi Damaghi, Zainab Raza, Rana Adil Farman, JiDong Bai, Joel Saltz, Bethany L. Niell. Physical tumor microenvironment biomarkers to predict DCIS progression, upstaging, and early metastasis [abstract]. In: Proceedings of the AACR Special Conference on Rethinking DCIS: An Opportunity for Prevention?; 2022 Sep 8-11; Philadelphia, PA. Philadelphia (PA): AACR; Can Prev Res 2022;15(12 Suppl_1): Abstract nr B018.
Abstract IA010: Patient-derived and genetically engineered models of Ductal Carcinoma in Situ
Authors: Jonkers Jos
Published: December 2022

Ductal Carcinoma in Situ (DCIS) is a non-invasive non-obligate precursor of invasive breast cancer (IBC). DCIS is usually treated by surgery combined with radiotherapy, which can have a large impact on the life of patients. However, many of these DCIS lesions would never progress into IBC. To reduce the overtreatment of DCIS, but assure proper treatment for high-risk DCIS, it is crucial to understand the biology underlying DCIS. Therefore, we established patient-derived xenograft (PDX) mouse models of DCIS by intraductally injecting patient DCIS material into the mammary ducts of female immunocompromised mice. In addition, we used somatic engineering approaches to establish genetically engineered rat models of DCIS. Together, these preclinical in vivo model platforms enable us to (i) identify biomarkers able to distinguish high-risk from low-risk DCIS, (ii) functionally validate candidate DCIS genes, (iii) study the contribution of tumor cell-intrinsic and -extrinsic processes to DCIS progression, and (iv) study the response to targeted therapeutics.

Citation Format: Jos Jonkers. Patient-derived and genetically engineered models of Ductal Carcinoma in Situ [abstract]. In: Proceedings of the AACR Special Conference on Rethinking DCIS: An Opportunity for Prevention?; 2022 Sep 8-11; Philadelphia, PA. Philadelphia (PA): AACR; Can Prev Res 2022;15(12 Suppl_1): Abstract nr IA010.
Abstract IA012: Mammary epithelial architecture modulates field cancerization
Authors: Messal Hendrik , Scheele Colinda , Lips Esther , Lutz Catrin , Hutten Stefan , Kristel Petra , Vlahu Carmen , Jonkers Jos , Wesseling Jelle , van Rheenen Jacco
Published: December 2022

The architecture of the human breast undergoes extensive remodeling throughout puberty, the menstrual cycle, pregnancy and involution with a risk of accumulating genetic alterations and tissue changes. Cells that have acquired mutations in driver genes are found to be abundant in tissues of healthy individuals, but they rarely develop into tumors. Yet, the underlying protection mechanisms that prevent tumor formation are largely unknown. Here, we utilized tissue clearing and imaged 15 normal human breasts and DCIS resections with cellular resolution to unravel the 3D architecture of thousands of mammary ducts. Surprisingly, our analysis revealed that disease-free breasts carry a high burden of microscopic tissue lesions. Distinct trees from the same breast differ in the kind and frequency of lesions, which coevolve scattered along the ductal routes. Interestingly, combining tissue clearing of FFPE archival material with spatial CNV-seq reveals that in half of the cases disease-free ductal segments neighboring DCIS lesions contain molecular alterations. To resolve the mechanisms that restrain and permit mutation spread, we compare the clonal fate behavior of cells that acquire oncogenic mutations with those bearing neutral mutations in the mouse mammary gland epithelium. From the quantitative analysis of the clonal dynamics, we find that local tissue remodeling during the estrous cycle leads to the stochastic elimination of the majority of mutant clones, while massively accelerating the expansion of a minority of clones that, by chance, survive. Thereby, oncogenic mutations can be spread along entire ducts before signs of transformation become visible. Together this data provides quantitative understanding of the earliest stages of epithelial evolution preceding disease.

Citation Format: Hendrik Messal, Colinda Scheele, Esther Lips, Catrin Lutz, Stefan Hutten, Petra Kristel, Carmen Vlahu, Jos Jonkers, Jelle Wesseling, Jacco van Rheenen. Mammary epithelial architecture modulates field cancerization [abstract]. In: Proceedings of the AACR Special Conference on Rethinking DCIS: An Opportunity for Prevention?; 2022 Sep 8-11; Philadelphia, PA. Philadelphia (PA): AACR; Can Prev Res 2022;15(12 Suppl_1): Abstract nr IA012.
Abstract IA016: The DCIS: A biological challenge and clinical dilemma
Authors: Srivastava Sudhir
Published: December 2022

Currently, one of the most challenging problems in oncology is predicting more precisely whether lesions that are detected by sensitive screening tests are indolent (hence, not requiring extensive treatment) or progressive and potentially life-threatening. For example, the increase in breast cancer “incidence” observed over the last 20 years is largely attributable to increased detection of ductal carcinoma in situ (DCIS) and stage I disease by imaging. Despite the increase in early-stage disease, the rates of advanced stage disease have changed only minimally, and the rate of metastatic disease has been virtually unchanged. Conversely, all screening tests miss some life-threatening cancers, known as “interval cancers,” that are diagnosed between scheduled screening tests. Breast cancer best illustrates the problem of interval cancers. It is known that more-aggressive types of breast cancer are more likely to be missed by screening; the cancer grows so quickly it becomes symptomatic after a normal screening test. For example, interval-detected breast cancers are 1.8 to 2.6-fold more likely to be estrogen receptor (ER) negative compared to screen-detected tumors, are likely to be diagnosed at a more advanced stage and are faster growing on average than screen-detected cancers. Relatively little is known about the molecular and cellular differences between screen- detected lesions and interval tumors which might be used to distinguish among the two types of lesions and potentially point to more effective strategies and targets for intervention. The National Cancer Institute has established programs to identify cellular and molecular characteristics that distinguish progressive from non-progressive lesions. Recent progress in this direction will be discussed by the speaker.

Citation Format: Sudhir Srivastava. The DCIS: A biological challenge and clinical dilemma [abstract]. In: Proceedings of the AACR Special Conference on Rethinking DCIS: An Opportunity for Prevention?; 2022 Sep 8-11; Philadelphia, PA. Philadelphia (PA): AACR; Can Prev Res 2022;15(12 Suppl_1): Abstract nr IA016.
Abstract A007: Assessing immunoprevention efficacy of an agonist of CD137 pathway in a tobacco carcinogen-induced lung cancer preclinical model
Authors: Gulen Ayse Ece , Tarique Mohammad , Ulker Vahap , Agca Evrim , Shirwan Haval , Yolcu Esma S.
Published: December 2022

Purpose: Lung cancer is the leading cause of cancer-related deaths worldwide. Cigarette smoking is the primary risk factor for lung cancer and is linked to 80-90% of lung cancer deaths according to the Centers for Disease Control (CDC). The most carcinogenic component of tobacco smoke is NNK (4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone). SA-4-1BBL is a novel CD137 receptor agonist that has shown cancer immunoprevention efficacy in transplantable preclinical tumor models by invoking an innate immune surveillance mechanism involving CD4+ CD44+ T and NK cells. As a preclinical model for high-risk populations, we evaluated the immunoprevention efficacy of SA-4-1BBL in lung tumor formation induced by NNK. Experimental Procedures: A/J female mice were used as a preclinical model for lung cancer immunoprevention due to their susceptibility to tobacco-carcinogen-induced lung cancer. A/J mice were i.p. injected with NNK weekly for eight consecutive treatments. The mice were also treated s.c. with 100 µg SA-4-1BBL twice, two weeks apart on weeks 6 and 8 of NNK treatment. Animals were monitored for 21 weeks and euthanized to harvest the lung and lung-draining lymph nodes for histology (H&E staining and PCNA immunofluorescence staining). In separate groups, mice were treated with a depleting antibody to CD4 twice two weeks apart on weeks 6 and 8 of NNK treatment to investigate the role of CD4+ immune cells in cancer immunoprevention. Results: NNK treatment resulted in lung tumor formation as assessed by PCNA immunofluorescence staining in all saline-injected control mice by the 21-week experimental end-point. Treatment with SA-4-1BBL resulted in significantly fewer macroscopic and microscopic tumor numbers than the control group (p < 0.01). Animals treated with SA-4-1BBL and subjected to CD4+ cell depletion showed similar numbers of macroscopic and microscopic lung tumors to the vehicle group. Conclusions: Prophylactic treatment with SA-4-1BBL significantly prevents NNK-induced lung tumor formation and CD4+ immune cells play a critical role in the observed immunoprevention efficacy of SA-4-1BBL. Supported by a grant from DOD LC190524 to Esma S. Yolcu.

Citation Format: Ayse Ece Gulen, Mohammad Tarique, Vahap Ulker, Evrim Agca, Haval Shirwan, Esma S. Yolcu. Assessing immunoprevention efficacy of an agonist of CD137 pathway in a tobacco carcinogen-induced lung cancer preclinical model [abstract]. In: Proceedings of the Second Biennial NCI Meeting: Translational Advances in Cancer Prevention Agent Development (TACPAD); 2022 Sep 7-9. Philadelphia (PA): AACR; Can Prev Res 2022;15(12 Suppl_2): Abstract nr A007.
Abstract A015: The β-blocker carvedilol prevents DMBA-induced mammary gland tumors
Authors: Sardar Pabitra K. , Shahid Ayaz , Yeung Steven , Andresen Bradley T. , Huang Ying
Published: December 2022

Breast cancer is the most diagnosed cancer globally and is the fifth leading cause of cancer related mortality worldwide. Chemoprevention is a promising method of reducing cancer mortality. However, there are no drugs that are used routinely as breast cancer preventative agents. Carvedilol (CAR), a beta-adrenergic receptor blocker, use in humans reduces breast cancer incidence; therefore, we hypothesized that carvedilol acts as a breast cancer preventative agent. Two protocols were used to evaluate the ability of carvedilol to prevent 15 mg 7,12-Dimethylbenz(a)anthracene (DMBA)-induced mammary tumors. In protocol 1, two doses of carvedilol (2 and 10 mg/kg) were provided as a 7-day pretreatment and throughout the study. In protocol 2, 10 mg/kg tamoxifen and 10 mg/kg carvedilol were provided as a 7-day pretreatment and treatment was halted after six weeks. In all studies carvedilol and tamoxifen were administered in the drinking water, animals were palpated weekly beginning at week 5 to detect the presence and location of mammary tumors, tumors were measured via calipers, and the experiments terminated at week 13. In protocol 1, 10 mg/kg, but not 2 mg/kg carvedilol was effective in delaying the tumor occurrence (P = 0.0002); the first appearance of tumors and median tumor appearance were separated by 6 and 4 weeks, respectively, compared to rats receiving DMBA alone. However, the tumors that appeared, grew at similar rate as those in the DMBA group. In protocol 2, tamoxifen prevented all but 1 rat developing a tumor and was similar to the negative control; however, 10 mg/kg carvedilol was indistinguishable from the DMBA group. Although tamoxifen prevented DMBA-induced breast cancer, the rats drank significantly less water resulting in statistically lower body weight than all other groups. The rats also drank significantly less of the carvedilol-laced water, but not to a degree that affected their body weight. Therefore, continuous carvedilol treatment is essential to achieve the observed chemopreventive effects.

Our data demonstrates the chemopreventive activity of carvedilol in a rat model of mammary gland carcinogenesis. The results provide significant implications in breast cancer chemoprevention using carvedilol, which is a safe FDA-approved medicine.

Citation Format: Pabitra K. Sardar, Ayaz Shahid, Steven Yeung, Bradley T. Andresen, Ying Huang. The β-blocker carvedilol prevents DMBA-induced mammary gland tumors [abstract]. In: Proceedings of the Second Biennial NCI Meeting: Translational Advances in Cancer Prevention Agent Development (TACPAD); 2022 Sep 7-9. Philadelphia (PA): AACR; Can Prev Res 2022;15(12 Suppl_2): Abstract nr A015.
Abstract A018: Targeting STAT3 for bladder cancer prevention – in vitro studies using spheroid and organoid models
Authors: Singh Surya P , Pathuri Gopal , Asch Adam , Cholewa Brian , Shoemaker Robert , Rao Chinthalapally V. , Madka Venkateshwar
Published: December 2022

Signal Transducer and Activator of Transcription 3 (STAT3) is tightly regulated in normal cells to maintain a transiently active state. In contrast, persistent STAT3 activation is frequently observed in bladder cancer (BC) and is associated with poor prognosis and chemoresistance. Hence, developing small molecule inhibitors targeting STAT3 may be helpful for preventing BC progression and improving the survival rate of patients with metastatic BC. Recently, the use of three-dimensional in vitro models in drug development has gained popularity as they closely resemble, to an extent, the in vivo environment in heterogeneity and physiological conditions.

Here we established spheroid and organoid models for bladder cancer and evaluated STAT3 inhibitors (C188-9 and SH5-07) for their anticancer activity in vitro.

Initially, we optimized the spheroid growth from human, rat, and mouse BC cell lines (J82, NBT-II, MB49, respectively) and tumoroid growth from the BBN-rat bladder cancer model. The anticancer efficacy of C188-9 and SH5-07 was evaluated in vitro at various doses (0-50 µM) in the 3D models of BC. Assays were performed to determine spheroid viability (calcein AM (CA) and EtBr staining), ATP and ROS production (MitoSOX™). Protein isolated from control and drug treated spheroids/tumoroids was used to evaluate pharmacodynamic biomarkers of cell proliferation, apoptosis, and STAT3 signaling. We demonstrate that treatment with C188-9 and SH5-07 significantly decreased the spheroids size (39-45% smaller compared to untreated, p<0.0001) along with decreased ATP (20%-40%, p<0.05), and pSTAT3 protein expression in spheroids derived from BC cell lines and rat BC organoids. Further, MitoSOX™ staining showed that STAT3 inhibitor treatment induced mitochondrial mediated ROS generation in BC spheroids. CA and EtBr staining showed that C188-9 and SH5-07 treatment induced cell death in BC spheroids that was also associated with caspase-3 cleavage.

These findings indicate that C188-9 and SH5-07 could suppress the activation of the STAT3 pathway and inhibit the bladder cancer spheroid growth by inducing ROS production and thus warrants further evaluation in vivo. Furthermore, our study provided valuable spheroid and organoid models for evaluating therapeutic candidates in an in vivo-mimic microenvironment, thereby providing great potential for drug testing. (Partly supported by P30CA225520 and Kerley-Cade Endowed Chair)

Citation Format: Surya P Singh, Gopal Pathuri, Adam Asch, Brian Cholewa, Robert Shoemaker, Chinthalapally V. Rao, Venkateshwar Madka. Targeting STAT3 for bladder cancer prevention – in vitro studies using spheroid and organoid models [abstract]. In: Proceedings of the Second Biennial NCI Meeting: Translational Advances in Cancer Prevention Agent Development (TACPAD); 2022 Sep 7-9. Philadelphia (PA): AACR; Can Prev Res 2022;15(12 Suppl_2): Abstract nr A018.
Abstract A019: Targeting STAT3 for lung cancer prevention
Authors: Siegfried Jill M. , Soloff Adam C. , Davis Autumn Gaither , Powers Amy A. , Eisenberg Seth H. , Stabile Laura P.
Published: December 2022

Former smokers have an elevated risk of lung cancer and account for a large proportion of newly diagnosed lung cancer. Former smokers with airway dysplasia have received modest benefit in randomized chemoprevention trials, while active smokers have not. More effective chemoprevention agents for former smokers at risk for lung cancer are needed. We previously reported that a cyclic double-stranded DNA oligonucleotide STAT3 decoy (CS3D) administered intravenously (IV) had strong anti-tumor effects in lung cancer xenograft models and was also effective as a chemoprevention agent using a tobacco carcinogen (NNK)-induced lung cancer mouse model that mimics “ex-smokers”. CS3D prevents the binding of activated STAT3 dimers to the promoter of target genes and induces p-STAT3 degradation. Our goal was to develop a clinically applicable direct delivery method of CS3D to the lungs. Using fluorescently tagged CS3D, we evaluated the feasibility and determined the optimal dosing schedule of CS3D administered intratracheally (IT, a mimic of human inhalation) compared to IV in mice. IT delivery 3 times/week provided a high initial drug level that was cleared from the lungs within a few hours, while IV dosing provided lower initial drug levels but persisted up to 8 hrs, with CS3D accumulation over time in the lungs. While the pharmacodynamic effect was also greater with systemic dosing, IT delivery produced inhibition of the STAT3 pathway. Phosphorylated STAT3, VEGF, IL6 and Ki67 expression in NNK-induced lung preneoplasias was down-modulated by IT CS3D delivery after 4 weeks of treatment compared to the mutant oligonucleotide (CS3M), confirming our previous findings with IV delivery. We also demonstrated that CS3D delivered IT altered the pulmonary environment by promoting a pro-inflammatory, anti-tumor response in myeloid cells of the lung directly, or by acting on tumor and stromal components to establish an immune-reactive tumor-microenvironment. No toxicities were found during IT or IV treatment and no organ abnormalities were detected by either regimen. A long-term chemoprevention study of CS3D delivered IT in the ex-smoker lung cancer murine model is underway. Our findings suggest that blocking STAT3 may be a useful strategy for lung cancer prevention, and may involve both inhibition of oncogenic signaling and enhanced anti-tumor immunity.

Citation Format: Jill M. Siegfried, Adam C. Soloff, Autumn Gaither Davis, Amy A. Powers, Seth H. Eisenberg, Laura P. Stabile. Targeting STAT3 for lung cancer prevention [abstract]. In: Proceedings of the Second Biennial NCI Meeting: Translational Advances in Cancer Prevention Agent Development (TACPAD); 2022 Sep 7-9. Philadelphia (PA): AACR; Can Prev Res 2022;15(12 Suppl_2): Abstract nr A019.
Abstract IA002: Combination of naproxen and a novel chemically-stable eicosapentaenoic acid analogue provide synergistic tumor protection in Pirc rats
Authors: Beach Ryan , Hatano Yuichiro , Qiao Yong , Grady James , Sei Shizuko , Mohammed Altaf , Rosenberg Daniel W.
Published: December 2022

Colorectal cancer (CRC) is the second leading cause of cancer-related deaths in the United States. Patients with the genetic disorder Familial Adenomatous Polyposis (FAP) develop hundreds to thousands of polyps that unless removed by prophylactic colectomy will progress to CRC at an early age. Non-steroidal anti-inflammatory drugs (NSAIDs) and ω-3 marine fish oils, such as eicosapentaenoic acid (EPA), have been evaluated for their chemopreventive potential in delaying the onset of CRC in high-risk patients. In this study, we determined whether the NSAID, naproxen, alone or in combination with a chemically-stable form of EPA (TP-252), affects tumor formation in the ApcPirc rat model. Rats fed a combination of naproxen and TP-252 exhibited a 95% reduction in tumor formation and a 98% reduction in tumor volume, respectively. To elucidate potential mechanisms of tumor protection, a comprehensive lipidomic analysis was performed on colonic mucosa to determine changes in eicosanoid metabolism. Animals receiving TP-252 alone or in combination with naproxen had significantly reduced mucosal levels of pro-inflammatory ω-6 eicosanoids (PGE2, 5-HETE, and 14,15-DiHETrE), along with a simultaneous increase in anti-inflammatory EPA-derived ω-3 eicosanoids. Our colonic mucosal lipidomic analysis also uncovered several potential pharmacodynamic (PD) lipid biomarkers, including resolvin E2, 9-HEPE, 12-HEPE and 18-HEPE, that were increased in both the tissue and plasma of rats receiving TP-252 and were significantly correlated with tumor protection. Further studies with this drug combination should be focused on dose optimization and the role of EPA-derived lipid mediators in CRC initiation and progression.

Citation Format: Ryan Beach, Yuichiro Hatano, Yong Qiao, James Grady, Shizuko Sei, Altaf Mohammed, Daniel W. Rosenberg. Combination of naproxen and a novel chemically-stable eicosapentaenoic acid analogue provide synergistic tumor protection in Pirc rats [abstract]. In: Proceedings of the Second Biennial NCI Meeting: Translational Advances in Cancer Prevention Agent Development (TACPAD); 2022 Sep 7-9. Philadelphia (PA): AACR; Can Prev Res 2022;15(12 Suppl_2): Abstract nr IA002.
Abstract IA005: Discovery and development of LFA-9 for CRC prevention
Authors: Rao Chinthalapally V.
Published: December 2022

Colorectal Cancer (CRC) is a major public health issue with an estimated 980,000 deaths annually, worldwide. Current trends show an increase of CRC incidence and mortality worldwide. Colonic adenomas are very common in ages 50 and over. Though, nonsteroidal antiinflammatory agents like Aspirin, Naproxen and COX-2 inhibitor and Celecoxib are useful to prevent the polyp progression and reducing CRC burden, continuous/chronic usage of these drugs are limited by GI toxicity and unwanted side effects. Thus, the rationale to establish safer anti-inflammatory agents for CRC prevention is important. Mechanistic studies suggest that up-regulation of 15-PGDH, sparing COX-1/2 and prostaglandin (PG)I2 synthase and/or selectively targeting mPGES-1 and 5-LOX would reduce the cardiovascular side effects and may improve the chemopreventive efficacy. There are limited to no studies on targeting mPGES-1/5LOX and it is one of the unexplored areas in colorectal cancer chemoprevention towards safer drug development. To design and develop selective inhibitors of dual mPGES-1/5-LOX, we used in-silico small molecular docking simulation approaches, and identified LFA-9 as a novel duel mPGES-1/5-LOX inhibitor among >35 analogs. Azoxymethane (AOM)-induced rat colonic tumors were utilized as ex-vivo to assess pharmacodynamic inhibitory effects of LFA-9 on mPGES-1 and 5-LOX by Radio-HPLC. In a series of animal experiments, we established the dose-range of toxicity and optimal doses of LFA-9. Potential preventive efficacy of LFA-9 was assessed AOM-rat colon carcinogenesis (Aberrant Crypt Foci, (ACF) as surrogate marker in male F344 rats and intestinal tumor inhibition in APCMin/+ mice. Furthermore, we have shown the efficacy of LFA-9 administered in the diet at the adenoma stage significantly suppressed colonic adenocarcinoma formation in AOM-induced colon cancer (sporadic CRC model) and PIRC rat model (FAP model). Overall, the above results show that LFA-9, a novel dual mPGES-1/5-LOX inhibitor, is a safer agent and has potential for prevention of CRC in high-risk patients. {This work was supported by NCI-PREVENT Cancer Program, R01 CA213987 and VA Merit Awards}

Citation Format: Chinthalapally V. Rao. Discovery and development of LFA-9 for CRC prevention [abstract]. In: Proceedings of the Second Biennial NCI Meeting: Translational Advances in Cancer Prevention Agent Development (TACPAD); 2022 Sep 7-9. Philadelphia (PA): AACR; Can Prev Res 2022;15(12 Suppl_2): Abstract nr IA005.
Abstract IA006: Reverse-engineering discovery of targets, involved cell types, and compounds for liver cancer chemoprevention
Authors: Hoshida Yujin
Published: December 2022

Hepatocellular carcinoma (HCC) is a cancer type with clinically unequivocal and well-defined predisposing conditions, namely chronic fibrotic liver diseases caused by viral hepatitis and metabolic disorders. By following up the at-risk patient cohort, natural history of HCC development can be captured over the timeframe of one to two decades. Archived biospecimens are often available from the time of initial liver disease diagnosis, and unbiased molecular profiling of the specimens provides unique opportunities to interrogate molecular dysregulations confidently correlated with the real long-term clinical outcome, i.e., HCC development from early-stage liver diseases. This “reverse-engineering” strategy helps increase the likelihood of identifying clinically relevant therapeutic targets without solely relying on data from experimental preclinical models. This is particularly useful in cancer chemoprevention target discovery given the the requirement of overwhelmingly lengthy and costly clinical validation of experimentally derived candidate targets, which is often practically infeasible. Utilizing this strategy, we could successfully identify HCC chemoprevention targets (e.g., cellular signaling such as EGFR pathway, circulating bioactive lipids), involved cell types (e.g., hepatic myofibroblasts, conventional dendritic cells), and compounds (e.g., small molecular signaling pathway inhibitors, generic agents), streamline the process of their clinical validation, and facilitate translation of promising chemoprevention agents to the care of liver disease patients.

Citation Format: Yujin Hoshida. Reverse-engineering discovery of targets, involved cell types, and compounds for liver cancer chemoprevention [abstract]. In: Proceedings of the Second Biennial NCI Meeting: Translational Advances in Cancer Prevention Agent Development (TACPAD); 2022 Sep 7-9. Philadelphia (PA): AACR; Can Prev Res 2022;15(12 Suppl_2): Abstract nr IA006.
Abstract IA008: How to move a preventative agent from design into approval
Authors: Krol Danielle
Published: December 2022

Cancer prevention is a global public health challenge that requires a multidisciplinary approach including biology, epidemiology, medicine, nutrition, education, clinical trials and public policy. An intent-to-treat analysis of a randomized controlled trial differs from that in the observational setting, therefore, it is important to understand the underlying disease process and aspects of the intervention through research. The genetic heterogeneity of some cancers strongly support an enhanced focus on prevention as a strategy in high-risk individuals. Important factors when developing preventative cancer agents include the time course of the intervention, dose and duration of exposure needed to effect risk reduction, trial endpoints, durability of the impact of intervention, and methodological problems in implementing and interpreting randomized trials to evaluate prevention strategies. The clinical development of cancer prevention therapeutics has many challenges, including the long duration necessary to conduct trials, the lack of adherence to therapy, and unanswered questions on the best endpoint remain. The government, whether it be local, state or national, plays a vital role in regulation and education. The history of chemoprevention regulatory approvals has taught us that these programs are important in the fight against cancer. The Food and Drug Administration has identified barriers to clinical trial participation and challenges in surrogate endpoints, although with a multidisciplinary approach including communication with industry, are extremely positive about the future direction of chemoprevention research.

Citation Format: Danielle Krol. How to move a preventative agent from design into approval [abstract]. In: Proceedings of the Second Biennial NCI Meeting: Translational Advances in Cancer Prevention Agent Development (TACPAD); 2022 Sep 7-9. Philadelphia (PA): AACR; Can Prev Res 2022;15(12 Suppl_2): Abstract nr IA008.
Abstract IA009: National Cancer Institute Small Business Innovation Research Development Center NCI SBIR & STTR resources for commercializing innovative cancer prevention
Authors: Franca-Koh Jonathan
Published: December 2022

The Small Business Innovation Research (SBIR) and Small Business Technology Transfer (STTR) Programs at the National Institutes of Health (NIH) serve as one of the largest sources of early-stage capital for technology commercialization in the United States (US). In FY2021, the National Cancer Institute (NCI) SBIR and STTR Programs provided approximately $182M in non-dilutive funding to US small businesses and managed over 400 different projects. By supporting startups and other small businesses in the cancer space, the NCI SBIR and STTR Programs act as catalysts of innovation for novel technologies and products to achieve NCI’s mission to prevent, diagnose, and treat cancer. Staffed by a dedicated team of scientists with wide-ranging technology and industry experience, the NCI SBIR Development Center provides both funding and non-funding resources to US small businesses all aimed at translating technologies to the clinic to help cancer patients and providers.

During the past 5 years, NCI has provided SBIR/STTR funding to approximately 25 companies who are developing novel cancer prevention agents. Companies focused on prevention of several indications such as colorectal cancer, oral squamous cell carcinomas, and human papillomavirus-related cancers. Award sizes ranged from $200k-$2M and included support for companies with technologies in the preclinical and clinical stages of development. NCI provided opportunities for non-funding support as well, including entrepreneurial training programs such as I-Corps™ at NIH and programs to facilitate connections between innovators and potential investors. Whether a technology is in the preclinical or clinical stage of development, NCI can help small businesses translate their innovative research to the marketplace so they become available to healthcare providers and their patients.

Citation Format: Jonathan Franca-Koh. National Cancer Institute Small Business Innovation Research Development Center NCI SBIR & STTR resources for commercializing innovative cancer prevention [abstract]. In: Proceedings of the Second Biennial NCI Meeting: Translational Advances in Cancer Prevention Agent Development (TACPAD); 2022 Sep 7-9. Philadelphia (PA): AACR; Can Prev Res 2022;15(12 Suppl_2): Abstract nr IA009.
Abstract IA010: NCATS Collaborative Translational Research/Drug Development Resources
Authors: Simeonov Anton
Published: December 2022

The mission of the National Center for Advancing Translational Sciences (NCATS) is to catalyze the generation of innovative methods and technologies that will enhance the development, testing, and implementation of interventions across a wide range of human diseases and conditions. NCATS is directly addressing this problem by discovering new technologies and other approaches that could greatly accelerate the process of developing and deploying therapeutic solutions. An introduction of the Center’s internal research operations will be presented, focusing on the use of advanced assay and chemistry technologies to address the wide range of needs associated with early discovery space. Further, an overview of collaborative opportunities for joint projects will be provided.

Citation Format: Anton Simeonov. NCATS Collaborative Translational Research/Drug Development Resources [abstract]. In: Proceedings of the Second Biennial NCI Meeting: Translational Advances in Cancer Prevention Agent Development (TACPAD); 2022 Sep 7-9. Philadelphia (PA): AACR; Can Prev Res 2022;15(12 Suppl_2): Abstract nr IA010.
Abstract IA011: Modulation of tumor associated macrophages in breast cancer
Authors: Guerriero Jennifer L.
Published: December 2022

Macrophages are an innate immune cell that play a critical role in host defense and maintaining tissue homeostasis, however their infiltration into tumors has been associated with disease progression and resistance to therapy. Tumor associated macrophages (TAMs) represent a significant proportion of solid tumors, including breast cancer. TAMs play a major role in tumorigenesis as they can enhance tumor cell growth, angiogenesis and metastasis. In addition, TAMs can inhibit anti-tumor responses of T cells. Our recent work has shown that removal or conversion of TAMs to an anti-tumor phenotype enhances chemo- and immuno-therapy establishing TAMs as targets for anti-cancer therapy. We recently revealed that a Class II HDAC inhibitor can polarize TAMs to an anti-tumor phenotype to facilitate reduction in primary and metastatic tumor burden. In addition, we have found that some types of therapy such as poly(ADP-ribose) polymerase (PARP) inhibition (PARPi) can drive development of highly suppressive TAMs, restricting anti-tumor T cell function and survival. Murine models demonstrate that in the absence of TAMs, PARPi induce a robust recruitment of cytotoxic T cells and durable antitumor responses. Therefore, targeting TAMs is a promising strategy for improving PARPi treatment efficacy. However, while there is an urgent need to target TAMs during tumorigenesis and cancer therapy, to date, failure to fully characterize TAM biology and classify multiple subsets has hindered advancement in therapeutic targeting. Deep analysis of TAMs in solid tumors has revealed the complexity of TAMs and revealed major gaps in our knowledge of the functional and phenotypic characterization of TAM subsets associated with cancer, before and after treatment. Here we will discuss the complexity of TAMs in solid tumors including characterizing TAM subsets, location, and crosstalk with neighboring cells, as well as novel TAM-modulating strategies and combinations that are likely to enhance current therapies and overcome chemo- and immuno-therapy resistance.

Citation Format: Jennifer L. Guerriero. Modulation of tumor associated macrophages in breast cancer [abstract]. In: Proceedings of the Second Biennial NCI Meeting: Translational Advances in Cancer Prevention Agent Development (TACPAD); 2022 Sep 7-9. Philadelphia (PA): AACR; Can Prev Res 2022;15(12 Suppl_2): Abstract nr IA011.