Profound Synergy Research Articles

Research on the Path of Aiding Integrating Excellent Traditional Chinese Culture into Biochemistry Teaching Curriculum with the Artificial Intelligence Technologies

Traditional culture has profound connotation and great inclusiveness. Incorporating traditional culture into biochemistry education not only boosts students' enthusiasm and attentiveness but also expands their analytical viewpoints. With the help of artificial intelligence technology, interweaving scientific and cultural wisdom, we can better promote our ancient traditions and instill a deep sense of national pride in our students. The article delves into the backdrop of merging Chinese exemplary traditional culture with the biochemistry curriculum. It further investigates the challenges of intensifying the instruction of this esteemed culture and the considerations when utilizing Chinese traditional values to enhance biochemistry education. On this basis, the article highlights the practical rationale for weaving Chinese exemplary traditional culture into the biochemistry curriculum, and briefly discusses the possible ways to improve the teaching effect of the course with the help of artificial intelligence technology, in order to promote the profound synergy between the three and thus promote the development of the biochemistry education framework.

Cell Context Is the Third Axis of Synergy for the Combination of ATR Inhibition and Cisplatin in Ewing Sarcoma.

The importance of cellular context to the synergy of DNA damage response (DDR)-targeted agents is important for tumors with mutations in DDR pathways, but less well-established for tumors driven by oncogenic transcription factors. In this study, we exploit the widespread transcriptional dysregulation of the EWS-FLI1 transcription factor to identify an effective DDR-targeted combination therapy for Ewing sarcoma. We used matrix drug screening to evaluate synergy between a DNA-PK inhibitor (M9831) or an ATR inhibitor (berzosertib) and chemotherapy. The combination of berzosertib and cisplatin was selected for broad synergy, mechanistically evaluated for Ewing sarcoma selectivity, and optimized for in vivo schedule. Berzosertib combined with cisplatin demonstrates profound synergy in multiple Ewing sarcoma cell lines at clinically achievable concentrations. The synergy is due to loss of expression of the ATR downstream target CHEK1, loss of cell-cycle check-points, and mitotic catastrophe. Consistent with the goals of the project, EWS-FLI1 drives the expression of CHEK1 and five other ATR pathway members. The loss of CHEK1 expression is not due to transcriptional repression and instead caused by degradation coupled with suppression of protein translation. The profound synergy is realized in vivo with a novel optimized schedule of this combination in subsets of Ewing sarcoma models, leading to durable complete responses in 50% of animals bearing two different Ewing sarcoma xenografts. These data exploit EWS-FLI1 driven alterations in cell context to broaden the therapeutic window of berzosertib and cisplatin to establish a promising combination therapy and a novel in vivo schedule. See related commentary by Ohmura and Grünewald, p. 3358.

Abstract 7123: Biomarker-targeted alkylator and DNA damage repair inhibitor combination for refractory and relapsed acute myeloid leukemia (AML)

Abstract Isocitrate dehydrogenase 1 and 2 (IDH1/2) convert isocitrate to α-ketoglutarate in the tricarboxylic acid (TCA) cycle. A missense mutation in IDH1/2 leads to accelerated conversion of α-ketoglutarate into 2-hydroxyglutarate (2-HG), an oncometabolite that competitively inhibits α-ketoglutarate-dependent dioxygenases including Ten-Eleven Translocation (TET) enzymes. TET enzymes are DNA demethylases that convert 5-methylcytosine to 5-carboxycytosine and regulate CpG island promoter methylation signatures. In IDH1/2 mutant glioma, inhibition of TET2 enzyme by 2-HG can lead to promoter hypermethylation of O6-methylguanine methyltransferase (MGMT) and reduce its expression. Approximately 10-15% of de novo or treatment-related AML harbor mutually exclusive mutations in IDH1/2 or TET2, and emerging resistance towards clinical IDH1/2 inhibitors, therefore warrants novel therapeutic options. We performed a pan-cancer TCGA analysis and observed MGMT promoter hypermethylation and reduced expression in IDH1/2 AML. Our independent study on 10 AML patients (5 IDH1/2 and 5 TET2 mutant) corroborated the TCGA data and showed a significantly reduced expression of MGMT in comparison to 7 wild-type AML samples. As MGMT resolves O6-alkylguanine lesions during DNA damage, we performed an exhaustive alkylator screen, where temozolomide (TMZ) and our novel compound KL-50 emerged as the top hits that imparted a maximum therapeutic index in multiple MGMT (MGMT+/–) isogenic AML cell line models. Both TMZ and KL-50 induced MGMT-dependent DNA strand breaks and replication stress and activated ATM and ATR DNA damage response pathways in a time-dependent manner. Inhibition of ATR activation with Ceralasertib, an ATR inhibitor (ATRi) currently in clinical trials for chronic myelomonocytic leukemia and myelodysplastic syndromes, imparted a profound MGMT-dependent synergy with TMZ and KL-50. Researchers have established in glioma that mismatch repair (MMR) loss confers resistance to TMZ, and reported studies also show evidence of MMR loss in refractory or relapsed AML patients. Thus, we developed numerous MMR-deficient isogenic AML cell lines in an MGMT+/– background and observed MMR loss-mediated resistance to TMZ and abrogation of its synergy with ATRi. Unlike TMZ, KL-50 retained a profound synergy with ATRi in an MGMT-dependent manner irrespective of MMR status. We are currently investigating our novel KL-50 and ATRi combination in MGMT/MMR isogenic cell line-based in vivo AML models and in AML patient-derived xenografts to establish it as a promising novel biomarker-targeted therapy for AML patients. Citation Format: Prateek Bhardwaj, Ranjini Sundaram, Amro Baassiri, Martin Matthews, Jennifer VanOudenhove, Susan Gueble, Stephanie Halene, Ranjit Bindra. Biomarker-targeted alkylator and DNA damage repair inhibitor combination for refractory and relapsed acute myeloid leukemia (AML) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7123.

Abstract 1082: Exploiting lurbinectedin-driven nucleolar relocalization of EWS-FLI1 to develop novel combination therapies for Ewing sarcoma

Abstract Background: Ewing sarcoma (ES) is a malignant bone tumor characterized by the oncogenic fusion protein EWS-FLI1. EWS-FLI1 acts as an aberrant transcription factor, inducing and silencing genes involved with increasing cell proliferation and survival. We have previously shown that lurbinectedin, a trabectedin analog, effectively inactivates the transcriptional activity of EWS-FLI1 by redistributing it within the nucleus to the nucleolus. Interestingly, this process activates a gene signature like that of the DNA damage response (DDR) to UV light, which was also shown to induce nucleolar translocation of WT Ewing sarcoma breakpoint 1 (EWSR1). However, the mechanism governing the nucleolar translocation of EWS-FLI1 induced by these drugs has yet to be elucidated. In addition, novel approaches to exploit this relocalization mechanism as a means to broaden the therapeutic window have yet to be established. Methods: We used immunoblotting, RT-qPCR, viability assays, cell fractionation, confocal microscopy, proximity ligation assays (PLA), mass spectrometry, genomics, and immunoprecipitation to investigate the mechanism of action of trabectedin/lurbinectedin-induced nucleolar relocalization of EWS-FLI1. Results: Through interrogation of the pathways triggered by trabectedin/lurbinectedin, we found that this mechanism is in part mediated by HSP70. Viability assays also revealed profound synergy between HSP70 inhibitors (HSP70is) and lurbinectedin. Immunoprecipitated EWS-FLI1 subjected to mass spectrometry revealed an association of EWS-FLI1 and several HSP70 isoforms after trabectedin treatment which was confirmed with PLA. Trabectedin/lurbinectedin treatment also revealed an upregulation of EWS-FLI1-suppressed targets and suppression of induced targets. Confocal microscopy showed that EWS-FLI1 “trapping” in the nucleolus was potentiated by combination treatment with HSP70is. Conclusion: Our results underscore the synergistic effect of combining HSP70is with trabectedin or lurbinectedin treatment to attenuate several aspects of ES tumorigenesis in vitro. In vivo experiments with patient-derived xenografts are underway to corroborate the synergy of HSP70is and trabectedin/lurbinectedin. Together, our data highlights the potential for a novel “EWS-FLI1 nucleolar trap” as a means to inhibit and sustain EWS-FLI1 repression with this class of compounds. Citation Format: Sridhar M. Veluvolu, Raphael D. Lopez, Jenna M. Gedminas, Elissa A. Boguslawski, Elizabeth R. Wilson, Benjamin P. Caiello, Maureen E. Murphy, Patrick J. Grohar. Exploiting lurbinectedin-driven nucleolar relocalization of EWS-FLI1 to develop novel combination therapies for Ewing sarcoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1082.

Artificial Intelligence and the Emergence of Co-Creativism in Contemporary Art

This paper argues for the emergence of a new art movement termed Co-Creativism, emblematic of the profound synergy between humans and artificial intelligence (AI) in shaping artistic narratives. Emerging as a successor to post-postmodernism and metamodernism, I propose Co-Creativism began its ascent around 2018 and has since solidified its prominence by 2023, notably influenced by the post-COVID landscape. The era transcends viewing AI as a mere instrumental entity, instead recognising it as an integral co-contributor in the creative realm. Through a methodical approach encompassing case studies and content analysis of artist statements, this paper aims to define the key characteristics and underlying themes of Co-Creativism. By examining the interplay between the global context, the art world, the notion of the artist, art-making practice, the audience, and co-creativist art, the goal is to provide a comprehensive understanding of Co-Creativism.

The Synergetic Paradigm: A Confluence of Linguistic and Cultural Anthropology in The Examination of Cultural Values

Cultural Anthropology and Linguistic Anthropology are two vital subfields within anthropology, each offering unique insights into the diverse and complex ways in which human societies operate. When these disciplines converge, a profound synergy emerges, providing a richer and more holistic understanding of the cultural, linguistic, and social dimensions of human life. This article explores the interplay between cultural and linguistic anthropology, revealing how their synergy enhances our grasp of the intricacies of human existence. The fields of cultural and linguistic anthropology have long been distinguished by their unique focuses, methodologies, and research interests. Cultural anthropology delves into the intricacies of human societies, exploring cultural practices, beliefs, and social structures, while linguistic anthropology focuses on the role of language in culture and society. However, a synergetic paradigm is emerging that seeks to bridge these two disciplines, recognizing the intricate connection between language and culture. This paradigm acknowledges that linguistic and cultural anthropology are not mutually exclusive but, instead, complementary lenses through which to view the complex tapestry of human societies. Cultural and linguistic anthropology, though distinct in their focuses, share a fundamental interest in understanding the dynamics of human societies and cultures. When it comes to exploring shifts in value systems, these two subfields of anthropology often employ similar research methods. In this article, we will delve into the methods that cultural and linguistic anthropologists commonly share in their investigations of evolving value systems. Keywords: Linguistic anthropology; Cultural anthropology; Shared methodology; values.

Phase 1/2 Study of Pacritinib in Combination with Azacitidine in Chronic Myelomonocytic Leukemia

Introduction and preclinical rationale Treatment options for chronic myelomonocytic leukemia (CMML) are limited. Hypomethylating agents (HMAs) are the only approved therapies but are associated with unsatisfactory response rates and do not impact disease evolution (Merlevede Nat Comm 2016). JAK inhibition can reduce GM-CSF signaling, which contributes to CMML pathobiology; ruxolitinib has been clinically evaluated in CMML demonstrating promising, but insufficient clinical activity. Pacritinib is a JAK inhibitor that has a unique profile which suggests it may be uniquely well suited for the treatment of CMML, including targeting of CSF1R and IRAK1, both of which are upregulated in CMML and related diseases (Obba Autophagy 2015, Rhyasen Cancer Cell 2013). Additionally, pacritinib was recently shown to be a potent inhibitor of ACVR1 (Oh AJH 2023), potentially allowing for increased iron availability and improvement in anemia. Pacritinib has been evaluated preclinically in CMML primary samples and mouse models where it demonstrated significant, albeit heterogenous activity (Yoshimi Blood 2017). To enhance the activity of pacritinib in CMML, it was also evaluated in combination with azacitidine in GM-CSF hypersensitive cell lines and primary CMML cells where profound synergy was noted (Ma ASH 2015). Based on encouraging preclinical data, we hypothesize that the combination of pacritinib with azacitidine will be an effective treatment strategy in CMML patients. Methods In this phase 1/2 trial, we will evaluate the safety and efficacy of pacritinib and azacitidine in newly diagnosed or previously treated CMML. Patients must be JAK inhibitor and HMA naïve (≤1 cycle of HMA allowed). Phase 1 will follow a ‘3+3‘ dose de-escalation design and will enroll 6-12 patients. The primary objective of this phase is to determine the recommended phase 2 dose (RP2D) of pacritinib in combination with a fixed dose of azacitidine. Pacritinib will be administered at 200mg twice daily (with a lower dose of 100mg twice daily being explored in the case of dose limiting toxicity) in combination with azacitidine 75mg/m 2 for 7 days. Phase 2 will utilize a Simon's two stage design to assess the preliminary efficacy of pacritinib in combination with azacitidine at the RP2D. In the first stage, after treating 6 patients at the RP2D, there will be an interim response assessment as part of the phase 2 portion. If at least 4 among these 6 patients show a response, the trial will proceed to a second stage and enroll an additional 12 patients. Patients will be treated for 6 cycles of 28 days each with an interim bone marrow biopsy after cycle 3. The primary endpoint of the phase 1 portion is the recommended RP2D through the assessment of dose limiting toxicities and of the phase 2 portion is the overall response rate (ORR) by the end of cycle 6 as defined by a clinical benefit or greater by myelodysplastic syndrome/myeloproliferative neoplasm International Working Group response criteria (Savona 2015). Secondary endpoints include duration of response, disease free survival, overall survival, changes in MPN-SAF total symptom scores, palpable splenomegaly, and adverse events. Several correlative studies are planned as part of this clinical trial. Modulation of biomarkers of disease activity with treatment including GM-CSF dependent STAT5 phosphorylation, MAPK, and PI3K signaling will be analyzed. We will also evaluate changes in mutational and cytogenetic burden with treatment as well as at baseline to identify predictive biomarkers of response.

Quantum Computing and AI: The Synergy of Two Revolutionary Technologies

An important turning point in the history of technology and computation is the confluence of Quantum Computing and Artificial Intelligence (AI). Redefining the limits of what is possible, quantum computing delivers previously unheard-of computational capabilities by utilizing the special qualities of quantum physics. AI, on the other hand, has made remarkable strides in simulating human intelligence, particularly through deep learning and natural language processing. This article explores the profound synergy arising from the intersection of Quantum Computing and AI. It explores the advantages and possible uses of this fusion, including how it can revolutionize the way complicated issues in drug development, cryptography, optimization, and other scientific fields are resolved. Additionally, it scrutinizes the challenges and ethical considerations inherent in this powerful merger. As Quantum Computing and AI continue to evolve and mature, their interplay promises to reshape industries and unlock new frontiers, bringing to life possibilities that were once confined to the realm of science fiction. This article navigates the exciting journey of these two groundbreaking technologies and their combined potential to revolutionize our world.

Visual design of green information in urban environment based on global similarity calculation and multi-dimensional visualization technology.

In recent years, the escalating prevalence of elevated consumption and carbon emissions within urban operations has reached a disconcerting extent. This surge in resource depletion and environmental pollution exerts an adverse influence on the well-being of individuals, while impeding societal progress and hindering the enhancement of overall quality of life. Within the domain of urban environmental design, the integration of visual displays emerges as a superior approach to facilitate the assimilation and analysis of green and low-carbon information. However, urban environmental data usually contains multiple dimensions, so it is a problem to realize the data representation of multiple dimensions while maintaining the correlation and interactivity between data. To surmount the challenge of visualizing such intricate information, this investigation initially employs a sophisticated memory-based clustering algorithm for information extraction, accompanied by a global similarity algorithm that meticulously computes attribute component quantities within specific dimensions of the vector. Furthermore, leveraging the inherent power of Vue's bidirectional data binding capabilities, the study adopts the esteemed MVVM (Model-View-View-Model) pattern, fostering seamless two-way interaction through the established logical relationship. As a result, the amalgamation of multidimensional visualization technology empowers comprehensive data mining through a captivating visual augmentation. Concurrently, the application of data visualization dimension control delivers tailored displays tailored to green and low-carbon scenarios within urban environmental design. Experimental results impeccably validate the effectiveness of the proposed algorithm, substantiated by a mere 1.77% false alarm rate for data stream difference detection and a clustering difference of 1.34%. The aforementioned algorithm accentuates the efficacy of visual displays, thus engendering a profound synergy between the industrial and supply chains. Moreover, it facilitates the design, production, and utilization of environmentally friendly products and energy sources. This, in turn, serves as a catalyst, propelling the widescale adoption of green and low-carbon practices throughout the entire industrial chain, fueled by the seamless integration of multimedia data.

BUSINESS ADMINISTRATION: "A DETAILED EXAMINATION OF LEADERSHIP STYLES AND THEIR INFLUENCE ON THE GROWTH AND SUCCESS OF START-UPS"

This paper provides a comprehensive review of leadership styles within the context of start-ups, exploring their effects on growth, innovation, and overall success. By carefully observing the delineation of leadership styles, including transformational, transactional, charismatic, situational, and laissez-faire, the review illuminates the unique attributes and implications each style holds for burgeoning businesses. Transformational leadership emerges as a crucial driver for innovation, emphasizing vision and collective drive. In contrast, transactional leadership offers operational efficiency but may risk long-term adaptability. Charismatic leaders, with their compelling personas, drive rapid growth, but centralization can pose challenges. With its inherent adaptability, situational leadership proves invaluable in the volatile start-up landscape, and laissez-faire leadership, promoting autonomy, demands careful balance to prevent directionlessness. The intricate relationship between leadership styles and business growth is explored in depth, underscoring that while leadership is a pivotal determinant, its efficacy intertwines with numerous variables, from market dynamics to team composition. One standout realization is the profound synergy between leadership and innovation, a lifeline for differentiation in the competitive start-up arena. Leadership’s role is thus not confined to mere administration; it shapes organizational culture, guiding innovation and strategic direction. The review emphasizes that effective start-up leadership is multifaceted, requiring foresight, adaptability, and a visionary outlook. As the entrepreneurial world continues to evolve, understanding and harnessing the nuances of leadership styles remains critical for sustained growth and success.

Understanding the nature of Pt-Rh synergy for three-way conversion catalysis

A series of Al2O3 supported Pt, Rh and Pt-Rh catalysts was investigated for TWC catalysis. Profound synergy between Pt and Rh was found on all Pt-Rh compositions relative to their single metal references with significantly reduced light-off temperatures. The nature of this synergy was probed using various characterization techniques. CO chemisorption, CO-DRIFTS and XPS results show that the surface Rh concentrations on aged Pt-Rh catalysts are much higher than that of the Rh reference. In addition, on the bimetallic catalysts the Rh species is in a more reduced state as indicated by XPS and XANES. AC-STEM/EDS results clearly illustrate that there are two types of Rh species on an aged Pt-Rh catalyst in terms of its location: one sitting on Pt particles as Rh nanoparticles and the other associated with the Al2O3 support as sub-nano Rh moieties. The Pt-Rh bimetallic particles contain a certain degree of Pt-Rh alloy as evidenced by the upshift of the metal XRD peaks and the Pt-Rh coordination from the EXAFS measurement. All structural and microscopy analyses suggest that Rh is surface enriched on the Pt-Rh particles, and it is the surface enriched Rh species that plays a primary role in enhancing the activity of Pt-Rh catalysts.

Highly Deformable and Durable Hydrogels through Synergy of Covalent Crosslinks and Nanosheet‐Reinforced Dynamic Interactions toward Flexible Sensor

AbstractIntegration of both excellent mechanical performance and remarkable durability into composite hydrogel materials is highly demanded yet challenging. Herein, a synergistic strategy of combining covalent crosslinks with nanosheet‐reinforced dynamic interactions is developed for the design and fabrication of deformable and recoverable nanocomposite hydrogels. Layered double hydroxide nanosheets and a trace amount of covalent crosslinks are simultaneously incorporated into the network, providing profound synergy in mechanical enhancement and durability extension. The resultant materials displayed high deformability (stretchability >1100%, compressibility >90%), considerable springiness and resilience, rapid and excellent recovery over 2000 cycles of compression and retraction as well as a 100% hysteresis ratio after only 1 min during large stretching. Notably, these networks are also transparent and ionically conductive. To demonstrate the utility, a wearable strain sensor is constructed for sensing and detecting human motions (e.g., subtle movement of mouth‐opening and large motion of joint‐bending), exhibiting a linear response with high sensitivity (gauge factor as high as 7.66) over 100–300% strain. This line of research will inspire renewed interest and work into synergic exploitation of nanospecies‐reinforced interactions and covalent crosslinks into the design of high‐performance composite materials.

Inhibition of Unc-51 like Autophagy Activating Kinase 1 (ULK1) Is Highly Synergistic with Chemotherapy and Bcl2 Inhibition in Acute Myeloid Leukemia (AML)

IntroductionAcquired resistance to chemotherapy and targeted therapies in AML may be mediated through autophagy. Bcl2 inhibition has also been reported to induce autophagy1. Therapeutic autophagy inhibition has been challenging because of limited druggable targets and pathway redundancy. Our preclinical work confirmed that Atg7 inhibition potentiates chemotherapy effects in AML2. While Atg7 is an E1-like activating enzyme,ULK1 is an apical kinase in the autophagy cascade and the only S/T kinase in the core autophagy pathway that phosphorylates multiple targets (Atg13, Beclin1 and VPS34). ULK1 initiates autophagosome formation and maturation. With the recent development of ULK1 inhibitors we were enabled to investigate the effects of ULK1 inhibition in AML.MethodsFirst we confirmed that chemotherapy (cytarabine, Ara-C) and Bcl2 inhibitor ABT199 induce autophagy in AML cells by LC3 quantification. Next, we studied potential synergy of these agents with ULK1 inhibitor, SBI-0206965 (SBI) in inhibiting cell proliferation and inducing apoptosis. For further insights into the mechanisms of synergy, we conducted large-scale proteomics analyses by reverse phase protein array (RPPA) and mass cytometry (CyTOF).ResultsAs a single agent SBI showed activity against a wide range of AML cell lines (IC50: 250 nM to 3.5 uM) and stem and progenitor cell populations (CD34+/CD38-) in primary AML samples with FLT3-ITD or p53 mutations, while sparing normal CD34+/CD38- cells. Immunoblotting confirmed target inhibition (Fig 1a) and apoptosis induction (Fig 1b).Treatment with Ara-C and ABT199 induced autophagy, which was reversed by SBI (Fig 2). Confirming our hypothesis, SBI showed significant synergy with both Ara-C (CI: 0.52) and ABT199 (CI: 0.16). Additional synergy was seen with FLT3-ITD inhibitor Sorafenib and MEK inhibitor Trametinib.Consistent with previous reports that autophagy deficiency induces DNA damage due to loss of ROS scavenging capacity, downregulation of DNA repair proteins and impaired protein complex formation at the sites of DNA damage3, we found that ULK1 inhibition results in elevated ROS generation which could be reversed by the ROS inhibitor N-acetylcysteine (NAC) (Fig 3a). We also found induction of DNA damage (Fig 3b). Autophagy deficiency has been recently shown to regulate DNA repair via Chk1 downregulation4. Our RPPA data showed significant decrease in cell cycle regulators phosphoS807/811 Rb, CDK1 and Chk1 (Fig 3b). Cell cycle analysis by EdU incorporation assay showed G1 arrest upon treatment with SBI (Fig 3c). In contrast, Bcl2A1 (pro-survival Bcl2 family member) was the most upregulated protein after treatment with SBI; rationalizing our combination studies with ABT199, which exhibits profound synergy (CI: 0.16) associated with increased cleaved Caspase7 and DNA damage (g-H2AX) (Fig 3b).Combinatorial studies with Ara-C also showed increased DNA damage (Fig 4a). Finally, CyTOF confirmed decreased phosphorylation of important signaling molecules like Akt, Erk1/2, MEK1/2, JNK, p38) upon SBI treatment (Fig 4b).We are conducting confirmatory mechanistic studies (changes in DNA damage response, ROS generation, etc.) by genetic silencing of ULK1 as well as studying the effect of drug combinations in the presence or absence of ULK1 inhibition in in vivo models of AML.ConclusionULK1 inhibitor SBI-0206965 effectively reverses therapy induced autophagy and synergizes with chemotherapy and Bcl2 inhibition imparting anti-leukemic effects, thus establishing a therapeutic rationale for ULK1 inhibition in AML. We are pursuing studies to validate the role of ULK1 inhibition in leukemia establishment, engraftment and survival in mice.

In Memoriam: José Baselga's Journey in Cancer Medicine.

In Memoriam: José Baselga's Journey in Cancer Medicine.

Abstract LB-148: Combination treatment of the CHK1 inhibitor, SRA737, and low dose gemcitabine demonstrates profound synergy with anti-PDL1 inducing durable tumor regressions and modulating the immune microenvironment in small cell lung cancer

Abstract Background: Small cell lung cancer (SCLC), the most aggressive form of lung cancer, shows poor response rates to immunotherapy targeting the programmed cell death protein 1 pathway (PD-(L)1). Our group previously discovered that SCLC exhibits high expression of checkpoint kinase 1 (CHK1) and that the CHK1 inhibitor SRA737 activates the innate immune STING pathway, demonstrating robust anti-tumor activity and synergy in combination with anti-PD-L1 in an SCLC model. As SRA737 is being tested in SCLC patients in combination with low dose gemcitabine (LDG), we evaluated the efficacy and immune correlates (including macrophages associated with resistance to immune checkpoint blockade) of the SRA737+LDG regimen in combination with anti-PD-L1 in an SCLC model. Results: Trp53, Rb1 and p130 (RPP) triple knockout SCLC cells were implanted into the flank of B6129F1 immunocompetent mice. After the mice developed tumors, they were treated with single agents or various drug combinations. Anti-PD-L1 and LDG demonstrated minimal effect on tumor growth as single agents and only a modest effect as a combination. Moderate to strong anti-tumor activity was however observed with SRA737 monotherapy which directly correlated with dosing intensity. The most profound and synergistic anti-tumor activity was observed when anti-PD-L1 was combined with the SRA737+LDG regimen, with all animals showing durable regressions. Analysis of tumor infiltrating immune cells at the end of this treatment regimen showed a dramatic induction of cytotoxic T-cells and a reduction of exhausted and regulatory T cells. Similarly, pro-inflammatory M1 type macrophages and dendritic cells were increased while immunosuppressive M2 type macrophages and MDSC cells were dramatically decreased. As monotherapy, the more dose intensive SRA737 schedule resulted in similar effects on lymphocytes when combined with anti-PD-L1. These effects are consistent with our previous data showing that SRA737 treatment leads to an induction of STING and type I interferon signaling in tumors, which is associated with the establishment of an anti-tumor immune microenvironment. Discussion: Our findings suggest that the combination of anti-PD-L1 with the SRA737+LDG regimen may represent the optimal implementation of these agents, leading to a dramatic anti-tumor activity accompanied by the establishment of a strong anti-tumor immune microenvironment. Given that anti-PD-(L)1 drugs are approved but show limited efficacy in SCLC, our preclinical data provide a strong rationale for combining these agents with the SRA737+LDG regimen to enhance clinical response rates. Citation Format: Triparna Sen, Carminia M. Della Corte, Snezana Milutinovic, Lixia Diao, Robert J. Cardnell, Ryan J. Hansen, Bryan Strouse, Michael P. Hedrick, Christian Hassig, Jing Wang, Lauren A. Byers. Combination treatment of the CHK1 inhibitor, SRA737, and low dose gemcitabine demonstrates profound synergy with anti-PDL1 inducing durable tumor regressions and modulating the immune microenvironment in small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr LB-148.

Inhibitory functions of PD-L1 and PD-L2 in the regulation of anti-tumor immunity in murine tumor microenvironment.

Although a role of PD-L1 in the suppression of anti-tumor immunity and its value as a predictive biomarker has been suggested by various preclinical and clinical studies, theprecise mechanisms how PD-L1 and PD-L2, another ligand of PD-1, regulate anti-tumor immunity in the tumor microenvironment are yet to be fully explored. Here, we address this issue using PD-L1-deficient tumor cells, PD-L1-knockout (KO) mice, anti-PD-L1 monoclonal antibody (mAb), and anti-PD-L2 mAb. Firstly, PD-L1-deficient or competent tumor cells were inoculated into wild-type or PD-L1-KO mice. Results of tumor growth and mouse survival indicated that both tumor- and host-derived PD-L1 are functional to suppress anti-tumor immunity, while the former contributes predominantly than the latter. Experiments using bone marrow (BM) chimeric mice, generated by transferring PD-L1-KO BM cells into wild-type mice or vice versa, further suggested that PD-L1 expressed on BM-derived hematopoietic cells mediates the suppressive effects on anti-tumor immunity. Secondly, anti-PD-L2 mAb treatment demonstrated a profound synergy with anti-PD-L1 mAb therapy, whereas anti-PD-L2 mAb alone hardly induced any anti-tumor effects, suggesting that PD-L2's function becomes evident when the effects of PD-L1 are abrogated by anti-PD-L1 mAb. Consistent with this notion, PD-L2 expression was upregulated on tumor-associated macrophages (TAM) when mice were treated with anti-PD-L1 mAb. Taken together, our study elucidated the importance of PD-L1 associated with tumor cells and non-tumor host cells, particularly BM-derived hematopoietic cells, as well as PD-L2 inducibly expressed on TAM in the suppression of anti-tumor immunity in the tumor microenvironment.

Lipopolysaccharide Potentiates PD-1 Targeted Therapies During Chronic Viral Infection

Programmed cell death-1 (PD-1) targeted therapies improve exhausted T cell responses during chronic infections and cancers. However, the efficacy of PD-1 blockade as a treatment to revert CD8 T cell exhaustion declines as the chronic infection progresses. Moreover, the majority of tumors do not respond to PD-1 blockade, highlighting an important clinical limitation. Recent reports have demonstrated an association between particular microbiota and the clinical efficacy of PD-1 targeted therapies, suggesting that certain bacterial components may influence the outcome of these therapies. In this study, we interrogated synergistic effects of bacterial lipopolysaccharide (LPS), a main component of Gram-negative bacteria and a strong inducer of TLR4 signaling. Using the model of chronic lymphocytic choriomeningitis virus (LCMV) infection in mice, we show that transient TLR4 stimulation with a low dose of LPS results in unprecedented potentiation of PD-1 targeted therapies via a dendritic cell intrinsic mechanism dependent on B7/CD28 costimulation. This profound synergy between PD-1 blockade and LPS was also observed in a model of advanced immune exhaustion that does not normally respond to PD-1 blockade. Taken together, our data demonstrate a potent synergy between LPS and PD-1 blockade at rejuvenating exhausted CD8 T cells during chronic infection, warranting the pre-clinical evaluation of TLR4 agonists to improve PD-1 targeted therapies.

WNT antagonists exhibit unique combinatorial antitumor activity with taxanes by potentiating mitotic cell death.

The WNT pathway mediates intercellular signaling that regulates cell fate in both normal development and cancer. It is widely appreciated that the WNT pathway is frequently dysregulated in human cancers through a variety of genetic and epigenetic mechanisms. Targets in the WNT pathway are being extensively pursued for the development of new anticancer therapies, and we have advanced two WNT antagonists for clinical development: vantictumab (anti-FZD) and ipafricept (FZD8-Fc). We examined the antitumor efficacy of these WNT antagonists in combination with various chemotherapies in a large set of patient-derived xenograft models. In responsive models, WNT blockade led to profound synergy with taxanes such as paclitaxel, and the combination activity with taxanes was consistently more effective than with other classes of chemotherapy. Taxane monotherapy increased the frequency of cells with active WNT signaling. This selection of WNT-active chemotherapy-resistant tumorigenic cells was prevented by WNT-antagonizing biologics and required sequential dosing of the WNT antagonist followed by the taxane. The WNT antagonists potentiated paclitaxel-mediated mitotic blockade and promoted widespread mitotic cell death. By blocking WNT/β-catenin signaling before mitotic blockade by paclitaxel, we found that this treatment effectively sensitizes cancer stem cells to taxanes. This combination strategy and treatment regimen has been incorporated into ongoing clinical testing for vantictumab and ipafricept.

Abstract SY43-02: Predicting solid tumor sensitivity to drug combinations using a multiplexed microinjection platform and prioritizing therapies for clinical development

Abstract Functional testing of Voruciclib combinations using the CIVO precision medicine platform. Joyoti Dey, Joey Casalini, Marc Grenley, Sally Ditzler, Angela Merrell, Jason Frazier, Matt Biery, Derek Thirstrup, Bill Kerwin, Emily Beirne, Connor Burns, Kate Gillespie, Tom Deckwerth, James Olson, Richard Klinghoffer Genomics-driven precision oncology has been exceptionally powerful in certain instances where therapeutically actionable genetic mutations drive cancer progression at the time of treatment and when redundant pathways or anti-apoptotic mechanisms fail to limit the efficacy of the selected therapeutic. Unfortunately, these cases represent a minority of solid tumors and many effective cancer drugs have pleiotropic effects that are not easily aligned with genomics-based efficacy predictions. In vitro functional analyses address this to a certain degree but in the absence of intact tumor microenvironment and the patient's immune system, both of which are important determinants of drug responses. The CIVO platform was designed to assess chemosensitivity, chemoresistance and underlying molecular mechanisms in a multiplexed fashion, in vivo, in human patients’ cancer or nonclinical models. The first human trial with CIVO was recently reported1 (Klinghoffer et al (2015). As our team works with the FDA to expand human precision medicine, we used the CIVO platform in canine patients and patient-derived murine models to establish translational strategies to advance investigational CDK inhibitor, Voruciclib in human clinical trials as well as to distinguish Voruciclib from other approved agents belonging to the same class of drugs. Using both our platform and conventional systemic administration, a direct comparison of Bortezomib in combination with Voruciclib or Palbociclib in a triple-negative breast cancer model revealed synergistic activity when Bortezomib was combined with Voruciclib, but not when combined with Palbociclib (see Poster/Abstract #6264). Further studies revealed that this effect is potentially linked to the MCL1-dependent basal subtype of TNBC. In addition to suppression of MCL1 activity and upregulation of Noxa activity, at a transcriptional level Voruciclib inhibits generation of the cytoprotective XBP1-spliced variant involved in the unfolded protein response (UPR) resulting in increased apoptosis as measured by cleaved PARP. The MCL1 suppression caused by Voruciclib in TNBC prompted us to evaluate Voruciclib as a single agent in DLBCL lymphoma and in combination with the BCL2 inhibitor, ABT199 (Venetoclax). In murine DLBCL models, Voruciclib and ABT199 showed modest activity when locally injected as single agents, but demonstrated profound synergy when injected in combination. To assess whether the synergy is restricted to cancer cells vs normal tissue and hence to assess the potential therapeutic window, we are currently assessing the systemic efficacy of the aforementioned combination using DLBCL tumor bearing mice. 1. Klinghoffer, R. A. et al. A technology platform to assess multiple cancer agents simultaneously within a patient's tumor. Sci Transl Med 7, 284ra258, doi:10.1126/scitranslmed.aaa7489 (2015). Citation Format: James Olson. Predicting solid tumor sensitivity to drug combinations using a multiplexed microinjection platform and prioritizing therapies for clinical development. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr SY43-02.

Pacritinib (PAC) Synergistically Potentiates Azacitidine (5AZA) Cytotoxicity in Chronic Myelomonocytic Leukemia (CMML)

CMML is an aggressive adult myeloid neoplasm hallmarked by peripheral monocytosis and a propensity for acute myeloid leukemia (AML) transformation. Although 5AZA and decitabine are FDA approved for use in CMML, no pharmacologic treatment has been shown to impact the natural history of this disease. We have previously demonstrated that the majority of CMML cases display GM-CSF hypersensitivity and that this pathway is a therapeutic target via GM-CSF neutralization or pharmacologic JAK2 inhibition. We have clinically tested the latter using the FDA approved JAK1/2 inhibitor ruxolitinib (RUX) with favorable results. Our data suggests that JAK inhibitors may be clinically effective and may serve as a foundation for future combination therapy in CMML. However, both RUX and 5AZA are associated with severe myelosuppression and therefore have overlapping toxicities that would make combination therapy clinically problematic. PAC is a JAK2 inhibitor that also inhibits CSF1 and IRAK1, also activated in CMML, and does not cause clinical myelosuppression. We therefore hypothesized that the combination of PAC and 5AZA may be an active and clinically feasible therapeutic combination in CMML.We first explored the preclinical activity of PAC in the monocytic and/or GM-CSF hypersensitive cell lines THP-1, U937, and MO7e (HL-60 negative control). These cell lines were cultured in the presence of GM-CSF (except for THP-1), and six increasing doses of PAC (0-10uM). After 48 hours, these cultures were analyzed for proliferation (MTT) and apoptosis (Annexin V/DAPI incorporation). In contrast to HL-60s, all monocytic cell lines tested had a dose dependent decrease in proliferation and increase in apoptosis with an IC50 approximating 0.5uM (Fig 1a). We additionally measured the capacity of PAC to augment GM-CSF dependent STAT5 phosphorylation in these cell lines by phos-flow. All cell lines had a dose-dependent decrease in pSTAT5 suggesting appropriate target inhibition. We next explored the activity of PAC in primary CMML BMNCs by measuring apoptosis and 14-day colony forming capacity (n=10). Using the exact dosing schema as above, we confirmed a dose dependent increase in apoptosis and decrease in colony forming capacity with an IC50 that approximated 0.5 uM (Fig 1b-c). When testing inhibition of basal and GM-CSF dependent STAT5 phosphorylation, PAC was able to inhibit pSTAT5 more potently than seen in cell lines.We next sought to determine whether PAC was synergistic in model cell lines by generating a synergy topology map using a proliferation assay that determines if there are synergistic doses of two drugs in an unbiased manner. We used the Bliss method to determine if formal synergy was present and identified profound synergy between PAC and 5AZA at a dose of approximately 0.5 and 2.5 uM respectively (fig 1d,e). Interestingly, the synergy topology of the PAC and 5AZA combination was distinct from that of RUX combination therapy suggesting a PAC-specific effect. We next confirmed our synergy results in primary CMML BMNCs by measuring the change in colony formation assay in control, PAC (0.5uM), 5AZA (2.5uM), or combination treated cells (n=5). As shown in Figure 1e, a decrease in colony forming capacity with combination therapy compared to other conditions was identified (p<0.001). However, no change in pSTAT5 inhibition was seen with combination therapy compared to other conditions suggesting an alternate mechanism of synergy.To explore the potential mechanisms responsible for the PAC and 5AZA synergy we performed whole transcriptome RNA seq of the above 3 model cell lines treated with PAC, 5AZA, RUX, and all possible combinations for 6 hours each to identify a PAC and 5AZA specific effect. Integrated RNA-seq analysis of drug-treated cell lines identified a unique expression signature associated with PAC and 5AZA that may provide insight into a mechanism of action explaining the synergy observed. Importantly, MTX2, SH3GLB1, and EIF5A2 were validated by qPCR to be dramatically increased in cells treated with the combination of PAC and 5AZA and are differentially expressed in AML. Functional studies depleting the above RNA targets are underway to determine if these transcripts are required for the synergy observed. Taken together, these data identified PAC as a preclinical therapeutic in CMML that is synergistic with 5AZA and may serve as a clinical feasible therapeutic strategy. [Display omitted] DisclosuresKomrokji:Celgene: Consultancy, Research Funding; Incite: Consultancy; Novartis: Speakers Bureau; GSK: Research Funding. List:Celgene Corporation: Honoraria, Research Funding. Padron:CTI: Honoraria, Research Funding; Incyte: Research Funding.