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Data on forest parameters defining the structure, health and condition of a forest stand is essential for forest management and conservation. The increasing frequency of forest changes, such as those caused by climate change-related drought and heat events, highlight the importance of having a forest database with high spatial and temporal resolution. Automated forest parameter extraction based on unmanned aerial vehicle (UAV) imagery is a cost-effective way to address the need for accurate and up-to-date forest data.The aim of this project is to develop user-friendly tools based on optical data from UAVs that can be applied to accurately and efficiently conduct digital forest inventories. We are using spectral and geometric information from UAV data to create methods for automated derivation of forest parameters such as diameter at breast height (DBH), tree stem positions, individual tree crown delineation, and coarse wood debris. These methods are being designed with the practical needs of potential users from the forestry sector in mind. Different flight configurations, such as nadir and oblique camera angles, as well as different acquisition times, were combined to generate structure from motion (SfM) data products (dense 3D point clouds, orthomosaics and height models) containing both ground and canopy information. For a study site within the Hainich National Park, Germany, we analyzed how leaf-off and leaf-on data can be combined to improve the derivation of stand parameters, such as tree stem positions and individual tree crowns, using point- and raster-based algorithms. Additionally, DBH on an individual tree basis was derived for the same study site using the cast shadows of tree trunks. To do so, a deep learning model was trained to identify stem shadows based on an orthomosaic of only ground points acquired during sunny and leaf-off conditions.

Glioblastoma patients have limited treatment options. Cancer stem-like cells (CSLC) contribute to glioblastoma invasiveness and repopulation; hence, they represent promising targets for novel therapies. BAL101553 is a prodrug of BAL27862, a novel microtubule-destabilizing agent inhibiting tumor cell proliferation through activation of the spindle assembly checkpoint, which is currently in phase I/II clinical development. Broad anticancer activity has been demonstrated against human cancer models, including tumors refractory to conventional treatments. We have shown that overexpression of microtubule + end-binding 1-protein (EB1) correlates with glioblastoma progression and poor survival. Here, we show that BAL27862 inhibits the growth of two glioblastoma CSLCs. As EB1 is overexpressed in the CSLC line GBM6, which displays a high tumorigenicity and infiltrative pattern of migration in vivo, we investigated drug activity on GBM6 according to EB1 expression. BAL27862 inhibited migration and colony formation at subcytotoxic concentrations in EB1-expressing control cells (GBM6-sh0) but only at cytotoxic concentrations in EB1-downregulated (GBM-shE1) cells. Three administrations of BAL101553 were sufficient to provoke an EB1-dependent survival benefit in tumor-bearing mice. Patterns of invasion and quantification of tumor cells in brain demonstrated that GBM6-sh0 cells were more invasive than GBM6-shEB1 cells, and that the antiproliferative and anti-invasive effects of BAL101553 were more potent in mice bearing control tumors than in EB1-downregulated tumors. This was associated with inhibition of stem cell properties in the GBM6-sh0 model. Finally, BAL27862 triggered astrocytic differentiation of GBM6 in an EB1-dependent manner. These results support the potential of BAL101553 for glioblastoma treatment, with EB1 expression as a predictive biomarker of response. Mol Cancer Ther; 15(11); 2740–9. ©2016 AACR.

Chemical structure of BAL27862 and BAL101553

Materials and methods: EB1 Western blot analysis GBM stem cells were lysed (lysis buffer: Tris 50 mM pH 8.0, NaCl 250 mM, Triton-X100 1%, SDS 0.1% with a cocktail of proteases and phosphatases inhibitors added freshly (all from Sigma-Aldrich). Thirty µg of total protein lysate were loaded onto a 12% SDS-PAGE gel. Nitrocellulose membrane (Bio-Rad laboratories, Marnes la Coquette, France) was blocked with 5% milk (powder) in Phosphate Buffer Saline (PBS) (Life technologies)-Tween (Sigma-Aldrich) pH 7.4 for 1 h and then incubated in PBS-Tween 0.1%-5% milk solution with mouse anti-EB1 antibody (clone 5, BD Biosciences, Le pont de Claix, France) (1/1000) and mouse α-tubulin (clone DM1A, Sigma Aldrich). After washing, membranes were incubated with anti-mouse peroxydase-conjugated secondary antibodies (Jackson Immunoresearch, Baltimore, USA) for 1 h. The bound antibodies were then detected using chemiluminescence detection kit (Millipore, Saint Quentin en Yvelines, France). Signals were recorded with G:BOX (Syngene/Ozyme, Saint Quentin en Yvelines, France ) and quantification was done with Image J software. Immunofluorescence analysis Indirect immunofluorescence was performed as previously described (15). Briefly, cells were fixed for 5 min with cold methanol (-20{degree sign}C), incubated with the anti-EB1 antibody 1/100 for 1 h and anti-mouse antibody Alexa 568 nm (Molecular Probes) 1/200; and FITC-coupled anti-α-tubulin antibody (clone DM1A; Sigma-Aldrich) 1/200 for 1 h at room temperature in the presence of DAPI for nuclear staining. For GFAP staining, cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton-X100 for 10 minutes. Primary antibody against GFAP (rabbit IgG, 10€‰Î¼g/ml, Dako) was incubated overnight at 4€‰{degree sign}C. Secondary antibody, Texas-red Goat anti-rabbit IgG (H+L) was purchased from Jackson Immunoresearch (Newmarket, UK), and was incubated for 1 h. Cells were analyzed using a Leica DM-IRBE microscope at a magnification of x100. Real-time quantitative PCR RNA samples were processed using a LightCycler 480 instrument (Roche Applied Science) and a LightCycler 480 SYBR Green I Master Mix (Roche Applied Science). Briefly, total DNA-free RNA (1 μg) was reverse-transcribed into cDNA using 1 μg of random hexamers and Superscript II reverse transcriptase as recommended by the manufacturer (Invitrogen Life Technologies, Cergy Pontoise, France). Measurements were performed in triplicate for each sample, and relative expression ratios of target gene transcripts (GFAP, TUBB3, CNP) and reference gene transcripts (18S, GAPDH, and ACTB) were calculated using qPCR efficiencies and cycle threshold (Ct) deviations of tumor and normal adult brain samples (control: Agilent Technologies) (25). RNA expression levels in GBM CSLCs were subsequently expressed as percentages compared to normal adult human brain samples corresponding to 100% of expression. Forward and reverse primers used for each gene are listed in Supplemental Table 1. Supplementary Table 1: Sequences of primers used in RT-qPCR Supplementary Figure 1: Chemical structure of BAL27862 and BAL101553 Supplementary Figure 2: Modulation of EB1 expression in GBM6 cells using siRNA or shRNA against EB1. (A) Western blot analysis of EB1 in GBM6 shEB1 clones (GBM6 shEB1, GBM6 shEB1#2), in GBM6 GFP shEB1 clone and in control clones (GBM6 wt, GBM6 sh0 and GBM6 GFP sh0). Ratios EB1/tubulin, relative to controls, from at least three independent experiments are presented under the blots. (B) Analysis of EB1 expression by Western blot of GBM6 wt or treated with siRNA against EB1 (GBM6 siEB1) or siRNA control (GBM6 si0). (C) Immunofluorescence staining of tubulin (green) and EB1 (red) in GBM6 wt, GBM6 si0 and GBM6 siEB1. Bar = 10 μm.

Sequences of primers used in RT-qPCR

Microtubules are major components of the cellular cytoskeleton, ubiquitously founded in all eukaryotic cells. They are involved in mitosis, cell motility, intracellular protein and organelle transport, and maintenance of cytoskeletal shape. Avanbulin (BAL27862) is a microtubule-targeted agent (MTA) that promotes tumor cell death by destabilization of microtubules. Due to its unique binding to the colchicine site of tubulin, differently from other MTAs, avanbulin has previously shown activity in solid tumor cell lines. Its prodrug, lisavanbulin (BAL101553), has shown early signs of clinical activity, especially in tumors with high EB1 expression. Here, we assessed the preclinical anti-tumor activity of avanbulin in diffuse large B cell lymphoma (DLBCL) and the pattern of expression of EB1 in DLBCL cell lines and clinical specimens.Avanbulin showed a potentin vitroanti-lymphoma activity, which was mainly cytotoxic with potent and rapid apoptosis induction. Median IC50 was around 10nM in both activated B cell like (ABC) and germinal center B cell like (GCB) DLBCL. Half of the cell lines tested showed an induction of apoptosis already in the first 24h of treatment, the other half in the first 48h. EB1 showed expression in DLBCL clinical specimens, opening the possibility for a cohort of patients that could potentially benefit from treatment with lisavanbulin.These data show the basis for further preclinical and clinical evaluation of lisavanbulin in the lymphoma field.

Ozonation is a powerful technique to remove micropollutants from wastewater. As chemical oxidation of wastewater comes with the formation of varying, possibly persistent and toxic by-products, post-treatment of the ozonated effluent is routinely suggested. This study explored an enzymatic treatment of ozonation products using the laccase from Trametes versicolor. A high-performance liquid chromatography coupled with high-resolution mass spectrometry (HPLC-HRMS) analysis revealed that the major by-products were effectively degraded by the enzymatic post-treatment. The enzymatic removal of the by-products reduced the ecotoxicity of the ozonation effluent, as monitored by the inhibition of Aliivibrio fischeri. The ecotoxicity was more effectively reduced by enzymatic post-oxidation at pH 7 than at the activity maximum of the laccase at pH 5. A mechanistic HPLC-HRMS and UV/Vis spectroscopic analysis revealed that acidic conditions favored rapid conversion of the phenolic by-products to dead-end products in the absence of nucleophiles. In contrast, the polymerization to harmless insoluble polymers was favored at neutral conditions. Hence, coupling ozonation with laccase-catalyzed post-oxidation at neutral conditions, which are present in wastewater effluents, is suggested as a new resource-efficient method to remove persistent micropollutants while excluding the emission of potentially harmful by-products. Graphical abstract

Background Glioblastoma (GBM) is an incurable disease with few approved therapeutic interventions. Radiation therapy (RT) and temozolomide (TMZ) remain the standards of care. The efficacy and optimal deployment schedule of the orally bioavailable small-molecule tumor checkpoint controller lisavanbulin alone, and in combination with, standards of care were assessed using a panel of IDH-wildtype GBM patient-derived xenografts. Methods Mice bearing intracranial tumors received lisavanbulin +/−RT +/−TMZ and followed for survival. Lisavanbulin concentrations in plasma and brain were determined by liquid chromatography with tandem mass spectrometry, while flow cytometry was used for cell cycle analysis. Results Lisavanbulin monotherapy showed significant benefit (P < .01) in 9 of 14 PDXs tested (median survival extension 9%-84%) and brain-to-plasma ratios of 1.3 and 1.6 at 2- and 6-hours postdose, respectively, validating previous data suggesting significant exposure in the brain. Prolonged lisavanbulin dosing from RT start until moribund was required for maximal benefit (GBM6: median survival lisavanbulin/RT 90 vs. RT alone 69 days, P = .0001; GBM150: lisavanbulin/RT 143 days vs. RT alone 73 days, P = .06). Similar observations were seen with RT/TMZ combinations (GBM39: RT/TMZ/lisavanbulin 502 days vs. RT/TMZ 249 days, P = .0001; GBM26: RT/TMZ/lisavanbulin 172 days vs. RT/TMZ 121 days, P = .04). Immunohistochemical analyses showed a significant increase in phospho-histone H3 with lisavanbulin treatment (P = .01). Conclusions Lisavanbulin demonstrated excellent brain penetration, significant extension of survival alone or in RT or RT/TMZ combinations, and was associated with mitotic arrest. These data provide a strong clinical rationale for testing lisavanbulin in combination with RT or RT/TMZ in GBM patients.

Body movements, from a short smile to a marathon run, are driven by muscle activity. Despite the fact that measuring muscle activity with electromyography (EMG) is technically well established, it is highly complex and its use in interfaces has been limited. Easy access to muscle sensing can offer new opportunities to Human-Computer Interaction (HCI) research. Off-the-shelf sensors often only provide low-level access, hence requiring expertise in signal processing and widening the gulf of execution for users without engineering skills. To address this challenge, we introduce EMBody, a data-centric toolkit for EMG-based interface prototyping and experimentation. EMBody offers multiple levels of prototyping fidelity for EMG sensing, signal processing, and data interpretation. Our data-centric toolkit encapsulates the different data representation stages, offering a wide range of customization opportunities to experts while also allowing non-technical designers to focus on creating new interaction techniques. EMBody features a lightweight form factor and wireless connectivity. Additionally, the system leverages an exploration-centered workflow by allowing rapid access to measurement data via the accompanying software. Users define a set of motions to be recognized and interactively provide example data points. The toolkit then handles signal processing and classification. The recognized movements are streamed on the local network, ready to be used by interactive applications. This paper reports on how to use EMBody and its implementation. We iteratively developed the toolkit in a series of workshops and example applications. Users who had none or very limited knowledge of EMG could rapidly create engaging functional prototypes, while experts appreciated the modularity of the software component allowing for a high degree of customization. We contribute the software and hardware components of EMBody as a tool for the research community to stimulate creative exploration of EMG systems.

Background Echocardiographic parameters of diastolic function depend on cardiac loading conditions, which are altered by positive pressure ventilation. The direct effects of positive end-expiratory pressure (PEEP) on cardiac diastolic function are unknown. Methods Twenty-five patients without apparent diastolic dysfunction undergoing coronary angiography were ventilated noninvasively at PEEPs of 0, 5, and 10 cmH2O (in randomized order). Echocardiographic diastolic assessment and pressure–volume-loop analysis from conductance catheters were compared. The time constant for pressure decay (τ) was modeled with exponential decay. End-diastolic and end-systolic pressure volume relationships (EDPVRs and ESPVRs, respectively) from temporary caval occlusion were analyzed with generalized linear mixed-effects and linear mixed models. Transmural pressures were calculated using esophageal balloons. Results τ values for intracavitary cardiac pressure increased with the PEEP (n = 25; no PEEP, 44 ± 5 ms; 5 cmH2O PEEP, 46 ± 6 ms; 10 cmH2O PEEP, 45 ± 6 ms; p n = 25): no PEEP, 0.76 ± 0.13 m/s; 5 cmH2O PEEP, 0.74 ± 0.14 m/s; 10 cmH2O PEEP, 0.68 ± 0.13 m/s; p = 0.016; peak A wave (n = 24): no PEEP, 0.74 ± 0.12 m/s; 5 cmH2O PEEP, 0.7 ± 0.11 m/s; 10 cmH2O PEEP, 0.67 ± 0.15 m/s; p = 0.014; E’ septal (n = 24): no PEEP, 0.085 ± 0.016 m/s; 5 cmH2O PEEP, 0.08 ± 0.013 m/s; 10 cmH2O PEEP, 0.075 ± 0.012 m/s; p = 0.002]. Conclusions PEEP does not affect active diastolic relaxation or passive ventricular filling properties. Dynamic echocardiographic filling parameters may reflect changing loading conditions rather than intrinsic diastolic function. PEEP may have slight positive inotropic effects. Clinical trial registration https://clinicaltrials.gov/ct2/show/NCT02267291, registered 17. October 2014. Graphical abstract

Introduction This review evaluates the clinical role of fibroblast growth factor receptor 2 (FGFR2) inhibition with derazantinib in patients with intrahepatic cholangiocarcinoma (iCCA) harboring actionable oncogenic FGFR2 fusions/rearrangements, mutations and amplifications. FGFR inhibitors such as derazantinib are currently being evaluated to address the unmet medical need of patients with previously treated, locally advanced or metastatic iCCA harboring such genetic aberrations. Areas covered We summarize the pharmacokinetics, and the emerging safety and efficacy data of the investigational FGFR inhibitor derazantinib. We discuss the future directions of this novel therapeutic agent for iCCA. Expert opinion Derazantinib is a potent FGFR1‒3 kinase inhibitor which also has activity against colony stimulating factor-1‒receptor (CSF1R) and vascular endothelial growth factor receptor‒2 (VEGFR2), suggesting a potentially differentiated role in the treatment of patients with iCCA. Derazantinib has shown clinically meaningful efficacy with durable objective responses, supporting the therapeutic potential of derazantinib in previously treated patients with iCCA harboring FGFR2 fusions/rearrangements. The clinical safety profile of derazantinib was well manageable and compared favorably to the FGFR inhibitor class, particularly with a low incidence of drug-related hand-foot syndrome, stomatitis, retinal and nail toxicity. These findings support the need for increased molecular profiling of cholangiocarcinoma patients.

Physical exercises can benefit our health, but avoiding improper form and overexertion is essential. Facilitating bodily insights can encourage learning about exercise form, allowing users to gain a deeper understanding of their physiology. To investigate this, we conducted a lab experiment where amateur users performed bicep curls, and interviews with sports coaches. Participants were provided with FitBack—a system that monitors muscle activity during exercises via electromyography (EMG) and offers real-time biofeedback. Amateurs reported that they were successful in improving their exercise form and could acquire deeper bodily insights. Coaches reflected on how understanding muscle activity through EMG could be effectively used for increasing body awareness during coaching, highlighting that EMG-based biofeedback is beneficial for a diverse set of users. Our work contributes insights into using bodily sensing to help users understand their bodies. We contribute guidelines for designing systems that use EMG biofeedback effectively in physical activity.

International audience; Micro- and nanomotors have seen substantial progress in recent years for biomedical applications. However, three grand challenges remain: (i) high velocities to overcome the blood flow, (ii) spatially selective control to enable complex navigation, and (iii) integration of a medical, tomographic real-time imaging method to acquire feedback information. Here, we report the combination of active magnetic matter and a medical imaging technique, namely magnetic particle imaging (MPI), which addresses these needs. We synthesize ∼200 nm magnetic nanoparticles and observe a macroscopic, collective effect in a homogeneous magnetic field with a rotating field vector. The nanoparticles form a millimeter-sized cloud and reach speeds of 8 mm s–1. This cloud is imaged and selectively steered with an MPI scanner. Our experimental results are supported by a model that highlights the role of the Mason number, the particle’s volume fraction, and the height of the cloud. The successful introduction of a fast swarm of microscopic units and the spatial selectivity of the technique suggest an effective approach to translate the use of micro- and nanobots into a clinical application.

Variations of the composition of cyanophycin (CGP) have been investigated since the early 2000s. Modifications of the polymer are of academical interest, but also expand the number of putative applications for CGP and its dipeptides in fields like food supplementations, and medical and cosmetic applications. Until recently variations of the composition occurred only in vivo. However, in the last years, two procedures using chemical or enzymatic in vitro modification were successfully applied. Since chemical treatments were more effective and reached higher conversion rates, a search for reagents and their applicability to conduct reactions with CGP was done. The reaction of CGP with methyl isocyanate resulted in the conversion of 50% of the lysine residues, while only 3% of the arginine residues were modified. However, using digested CGP dipeptides, the conversion rates of lysine increased slightly to 72%, while the conversion of arginine reached 96%. Using formaldehyde, CGP could be methylated with a conversion rate of 84% for lysine and 15% for arginine. Acetylation of lysine residues was obtained using acetic anhydride, reaching a conversion rate of 100% for a single acetylation, where 63% of the residues were acetylated twice. Arginine residues could be acetylated at a rate of 89%. Diacetyl could be added to 80% of all arginine residues, while lysine was not targeted by the compound. Other agents were also tested, but showed lesser or no conversion and/or inconclusive results. Overall, the tested reactions confirm the viability of chemical CGP modification for future approaches.

Background: BAL0891 is a dual inhibitor of threonine tyrosine kinase (TTK) and polo-like kinase 1 (PLK1). These kinases collaborate in activating the mitotic spindle assembly checkpoint (SAC) at the kinetochore (KT) to regulate chromosome alignment and segregation prior to mitotic exit. In vitro, BAL0891 has a combined prolonged effect on TTK and a transient effect on PLK1, leading to rapid disruption of the SAC that potentiates aberrant mitotic progression of tumor cells. In this work, efficacy of BAL0891 was investigated in mouse models of human triple negative breast cancer (TNBC) including evaluation of dose-dependency, drug exposure, target occupancy and a screen of activity across a panel of PDX models. Methods: The MDA-MB-231 cell line was grown sc in nude mice and treated with BAL0891, administered IV weekly (QW) or twice-weekly (2QW). Thirteen sc TNBC PDX models were screened for BAL0891 response using 2QW administration. Efficacy was quantified as deltaT/C (treated/control tumors). Plasma and tumor were analyzed for drug levels or TTK target occupancy by LC-MS/MS. The latter used a biotinylated TTK-specific probe and streptavidin-mediated isolation of unoccupied TTK, trypsin digestion and quantification of TTK-representative peptides. Results: BAL0891 efficacy was tested in the TNBC xenograft model MDA-MB-231 with QW or 2QW IV dosing schedules. All treatments were well tolerated, with no drug-related animal deaths. With MTD dosing, tumor regressions were observed, while different MTD fractions for both QW and 2QW schedules showed dose-dependent anti-tumor activity. The weekly MTD group was followed for an additional 20 days after treatment cessation on day 100. Strikingly, 3 of 8 tumors continued to shrink resulting in 2 (25%) pathologically confirmed cures. Consistent with the potent efficacy of intermittent MTD dosing, and prolonged tumor drug exposure, tumor TTK was fully drug-occupied for ≥ 6 days after the last administration; target occupancy was also dose-and drug exposure-dependent. To further evaluate BAL0891 anti-cancer activity in TNBC, a screen in 13 TNBC PDX models was conducted. Seven models exhibited deltaT/C < 50%, with regressions observed in 3. Of these, 2 models showed persistent regressions ≥ 70% vs. baseline. Interestingly, evaluation of TTK target occupancy in selected models showed high target occupancy independent of tumor response, indicating target dependency rather than drug availability is important for anti-cancer activity. Conclusion: BAL0891 is a novel dual TTK/PLK1 mitotic checkpoint inhibitor with potent anti-cancer activity in TNBC models. Intermittent IV administration is well tolerated and associated with prolonged tumor drug exposure, prolonged TTK inhibition and notable anti-tumor efficacy. These data support further investigation of BAL0891 for the treatment of cancer patients (incl. TNBC). Citation Format: Heidi A. Lane, Felix Bachmann, Elisa Zanini, Paul McSheehy, Karine Litherland, Nicole Forster-Gross, Luc Bury, Diep Vu-Pham, Jos de Man, Wilhelmina E. van Riel, Guido JR Zaman, Rogier C. Buijsman, Laurenz Kellenberger. BAL0891: A novel dual TTK/PLK1 mitotic checkpoint inhibitor (MCI) that drives aberrant tumor cell division resulting in potent anti-cancer activity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5645.

Background: BAL0891 is a dual inhibitor of threonine tyrosine kinase (TTK) and polo-like kinase 1 (PLK1). These kinases collaborate in activating the mitotic spindle assembly checkpoint (SAC) at the kinetochore (KT) to regulate chromosome alignment and segregation prior to mitotic exit. In this work, kinase inhibition by BAL0891 was linked to effects on SAC integrity and aberrant mitotic progression in tumor cells. Comparison with a TTK-specific inhibitor (CFI-402257, CFI) allowed further evaluation of the contribution of dual TTK/PLK1 inhibition to anti-cancer activity, associated with a promising anti-proliferative profile across diverse tumor cell lines. Methods: Kinase assays used a radiometric assay. Target residency was measured using surface plasmon resonance with recombinant kinase. Anti-proliferative activity was assessed with crystal violet or YO-PRO assay (5 days incubation), in-cell target inhibition by immunoblotting for phospho-TTK following drug wash-out. Effects on SAC integrity were followed by immunoprecipitation (IP) mitotic progression by flow cytometry/mitotic marker expression. Cells were blocked in mitosis using the microtubule-targeting agent nocodazole or the PLK1 inhibitor onvansertib. SAC KT accumulation was evaluated by immunofluorescence (IF) for co-localization of BubR1 with CENPC. Comparative studies with CFI used anti-proliferative IC50 concentrations. Results: In vitro kinase profiling showed that BAL0891 has low nM IC50s against TTK and PLK1, with prolonged TTK (>12 h) and transient PLK1 (4 min) target residency. Prolonged TTK inhibition (≥38 h) was also observed in HT29 tumor cells. Consistent with a dominant TTK-targeting activity, BAL0891 treatment of HT29 cells blocked in mitosis with nocodazole or the PLK1 inhibitor onvansertib led to aberrant mitotic release and accumulation of polyploid cells. This was preceded by SAC disruption as visualized by IP assays. Effects on the SAC and mitotic exit were evaluated in comparative studies with CFI; BAL0891 exhibited faster kinetics for both parameters suggesting a contribution of PLK1 inhibition. This was confirmed by directly evaluating acute effects on SAC integrity at the KT by IF. Specifically, 1 h BAL0891 treatment of mitotic HT29 cells resulted in a highly reproducible and significant reduction in KT-associated SAC (p5 uM). Conclusion: BAL0891 is a novel dual TTK/PLK1 mitotic checkpoint inhibitor. In tumor cells, prolonged effects on TTK and transient effects on PLK1 contribute to rapid SAC disruption and aberrant mitotic exit. This is associated with potent anti-proliferative activity in diverse tumor lines. Citation Format: Elisa Zanini, Nicole Forster-Gross, Felix Bachmann, Nicole Willemsen-Seegers, Jos de Man, Guido J. Zaman, Rogier C. Buijsman, Anna Groner, Mila Roceri, Karin Burger, Paul McSheehy, Laurenz Kellenberger, Heidi A. Lane. BAL0891: A novel, small molecule, dual TTK/PLK1 mitotic checkpoint inhibitor (MCI) that drives aberrant tumor cell division [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5646.

Background: Avanbulin (BAL27862) is a microtubule-targeting agent (MTA) that promotes tumor cell death by modulating the spindle assembly checkpoint. Preclinical studies in glioblastoma (GBM) have shown that avanbulin is also active in human cancer cells resistant to other MTAs used in the clinic. Lisavanbulin (BAL101553) is a water-soluble, oral, lysine prodrug of avanbulin, currently in phase 1/2 for patients with recurrent GBM and, in combination with radiotherapy, for newly diagnosed GBM patients. Preclinical studies and clinical signals have also suggested an association between lisavanbulin activity and expression of microtubule plus-end binding protein (EB1). MTAs are active agents for lymphoma patients, as exemplified by the inclusion of vincristine in the R-CHOP regimen, standard treatment for diffuse large B cell lymphoma (DLBCL). Thus, our aim was to evaluate avanbulin, the active moiety of lisavanbulin, for its potential anti-lymphoma activity. Methods: Lymphoma cell lines derived from activated B cell like (ABC) and germinal center B cell type (GCB) DLBCL were exposed to increasing doses of avanbulin; cell proliferation was measured with MTT. Apoptosis induction was performed with annexin V staining on cells incubated in a live cell imaging system (Incucyte) treated with 5, 10, 20 or 40nM avanbulin. Images were taken every 4h for 72h. Cell cycle analysis was performed after 24, 48 and 72h of drug exposure at 20nM. Results: 26 DLBCL cell lines, treated for 72h with increasing avanbulin doses, showed high anti-proliferative activity of the compound, with a median IC50 = 11nM. No differences in terms of IC50 were observed comparing ABC (n=8) to GCB (n=18) DLBCLs, nor based on MYD88, TP53 and BCL2 status. Cell lines with MYC translocation were less sensitive than the ones without translocation. All lymphoma cell lines exhibited high EB1 RNA expression, with no correlation with sensitivity to avanbulin.Live cell imaging demonstrated a strong apoptosis induction in 15 out of 17 cell lines tested at 20 and 40nM. Among the 15 cell lines undergoing apoptosis, 8 cell lines already showed an apoptosis induction within 24h of drug exposure, 6 lines between 24 and 48h and 1 line after 48h exposure. These data confirmed the high activity of avanbulin already seen with the MTT assay.Cell cycle experiments performed at 24, 48 and 72h confirmed the strong cytotoxic effect of avanbulin in 4 DLBCL cell lines, including 2 ABC (OCILY3 and TMD8) and 2 GCB (SUDHL5 and SUDHL16) lines. Sub-G0 accumulation was already present at 24h in all 4 cell lines tested, with an increase over time. Especially at 24h, where we also observed a G2-M arrest in 3 out of 4 cell lines. Conclusions: Our data demonstrate the high cytotoxic anti-lymphoma activity of avanbulin, suggesting a potential activity of its prodrug lisavanbulin in lymphoma patients. Citation Format: Filippo Spriano, Luca Aresu, Luciano Cascione, Alberto J. Arribas, Nicole Forster-Gross, Felix Bachmann, Heidi Lane, Francesco Bertoni. Avanbulin, the active moiety of the tumor checkpoint controller lisavanbulin (BAL101553), has anti-lymphoma activity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2575.

Microswimmers are smart devices with potential applications in medicine and biotechnology at the micrometer-scale. Magnetic micropropellers with their remote control via rotating magnetic fields are especially auspicious. Helicoidal propellers with a linear velocity-frequency dependence emerged as the standard propulsion mechanism over the last decade. However, with their functions becoming more pivotal on the way to practical uses, deviations in shape and swimming behavior are inevitable. Consequently, propellers with nonlinear velocity-frequency relationships arise that not only pose different challenges but also offer advanced possibilities. The most critical nonlinearities are the wobbling behavior with its solution branching that has potential for bimodal swimming and the swimming characteristics in the step-out regime that are essential for selection and swarm control. Here, we show experimentally and with numerical calculations how the previously unpredictable branching can be controlled and, thus, becomes utilizable with an example 3D-printed swimmer device. Additionally, we report how two step-out modes arise for propellers with a nonlinear velocity-frequency dependence that have the potential to accelerate future microswimmer sorting procedures., Max Planck Society; Deutsche Forschungs-gemeinschaft; Priority Program 1726 Microswimmers