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Supplemental Figure 2 data file

PARP1 IHC staining. (a) PARP1 staining of frontal cortex autopsy tissue of a diffuse midline glioma patient. Only a subset of cells show PARP1 staining, while others are negative. Neurons show staining of the nucleoli, while cells of oligodendritic/astrocyte nature show PARP1 expression in the entire nucleus. Other cells, inclusing endothelial cells, show no PARP1 expression. The corresponding H&E is from the same area as the PARP1 stained image. (b) Physiological, non-tumor related PARP1 expression was observed in the brain of ntv-a;p53fl/fl mice in the Purkinje cell layer in the cerebellum and the dentate gyrus. Other brain areas showed low levels of PARP1 expression. (c) Example of PARP1 and H&E staining of tumor bearing ntv-a;p53fl/fl mice injected with DF1 cells at 4-6 weeks of age.

SUMMARY Objective To compare the efficacy, color stability, and tooth sensitivity (TS) of combined bleaching, using a modified protocol with at-home bleaching. Methods Eighty participants were randomized into two groups. In the combined group, a desensitizing gel was applied (10 minutes) prior to in-office bleaching (35% hydrogen peroxide (H2O2), 2×15 minute applications) and at-home bleaching (4% H2O2, 2×30 minutes for 21 days) started the next day. In the at-home group, only the at-home bleaching was performed. Color was recorded at the beginning and postbleaching with two scales (VITA Classical and Bleachedguide) and Easyshade spectrophotometer. The TS was recorded daily with a 0–10 visual analogue scale (VAS) and five-point numeric rating scale (NRS). Results A 40% lower risk (RR=1.4; 95% CI 1.1–1.9) was observed in the at-home group. Higher color change and intensity of TS [mean difference 2.3 (95% CI 1.3–3.3) in the VAS] was observed in the first week for the combined group. After the end of the protocol, a bleaching degree was detected for both groups, with no significant difference between both groups (p>0.05). Conclusion The combined group produced a slightly higher degree of color change than at-home bleaching but with a higher risk and intensity of TS.

αv β6 Integrin is an epithelial transmembrane protein that recognizes latency-associated peptide (LAP) and primarily activates transforming growth factor beta (TGF-β). It is overexpressed in carcinomas (most notably, pancreatic) and other conditions associated with αv β6 integrin-dependent TGF-β dysregulation, such as fibrosis. We have designed a trimeric Ga-68-labeled TRAP conjugate of the αv β6 -specific cyclic pentapeptide SDM17 (cyclo[RGD-Chg-E]-CONH2 ) to enhance αv β6 integrin affinity as well as target-specific in-vivo uptake. Ga-68-TRAP(SDM17)3 showed a 28-fold higher αv β6 affinity than the corresponding monomer Ga-68-NOTA-SDM17 (IC50 of 0.26 vs. 7.4 nM, respectively), a 13-fold higher IC50 -based selectivity over the related integrin αv β8 (factors of 662 vs. 49), and a threefold higher tumor uptake (2.1 vs. 0.66 %ID/g) in biodistribution experiments with H2009 tumor-bearing SCID mice. The remarkably high tumor/organ ratios (tumor-to-blood 11.2; -to-liver 8.7; -to-pancreas 29.7) enabled high-contrast tumor delineation in PET images. We conclude that Ga-68-TRAP(SDM17)3 holds promise for improved clinical PET diagnostics of carcinomas and fibrosis.

Indolo[2,3-d]benzazepines (indololatonduines) are rarely discussed in the literature. In this project, we prepared a series of novel indololatonduine derivatives and their RuII and OsII complexes and investigated their microtubule-targeting properties in comparison with paclitaxel and colchicine. Compounds were fully characterized by spectroscopic techniques (1H NMR and UV–vis), ESI mass-spectrometry, and X-ray crystallography, and their purity was confirmed by elemental analysis. The stabilities of the compounds in DMSO and water were confirmed by 1H and 13C NMR and UV–vis spectroscopy. Novel indololatonduines demonstrated anticancer activity in vitro in a low micromolar concentration range, while their coordination to metal centers resulted in a decrease of cytotoxicity. The preliminary in vivo activity of the RuII complex was investigated. Fluorescence staining and in vitro tubulin polymerization assays revealed the prepared compounds to have excellent microtubule-destabilizing activities, even more potent than the well-known microtubule-destabilizing agent colchicine., Several synthesized indololatonduine derivatives and their RuII and OsII complexes were investigated for their microtubule-targeting properties in comparison with paclitaxel and colchicine. Fluorescence staining and in vitro tubulin polymerization assays indicate excellent microtubule-destabilizing activity. The compounds were even more potent than the well-known microtubule-destabilizing agent colchicine.

International audience; Here we present the methodology and results of transferring UV-NIR flux calibration from NIST photodiodes to a set of 20 picoammeters. These are to be deployed as flux reference sensors on the SCALA calibration system at the University of Hawaii 2.2m telescope on Maunakea as part of a systematic upgrade aimed at improving the existing flux calibration for dark energy and exoplanet host star measurements beyond the ∼ 4 mmag / 100 nm we have already achieved at optical wavelengths with SCALA. Our robotic light source for performing the photodiode calibration transfer provides monochromatic light spanning 230 to 1200 nm with a dynamic range of 106 , while our new picoammeters have a noise floor of 10 fA in 4 s at 25 °C, with saturation around 400 pA. Our robotic gantry enabled the measurement of the spatial and angular response of our picoammeters. In preparation for the calibration transfer, a number of tests were performed to establish the measurement uncertainties, and these tests revealed subtle systematic effects that required correction. These includes polarization effects, leading to the redesign of part of the optics in the gantry head, implementation of a Holmium-Didymium filter as a precision wavelength transfer between arc and continuum light sources, and further suppression of stray light. We find that our calibration transfers are consistent with the NIST calibration to within ∼0.1%

P. ostreatus biomass could stand as an alternative vegan protein source due to its high protein content and amino acid composition.

Deubiquitylases (DUBs) are therapeutically amenable components of the ubiquitin machinery that stabilize substrate proteins. Their inhibition can destabilize oncoproteins that may otherwise be undruggable. Here, we screened for DUB vulnerabilities in multiple myeloma, an incurable malignancy with dependency on the ubiquitin proteasome system and identified OTUD6B as an oncogene that drives the G1/S-transition. LIN28B, a suppressor of microRNA biogenesis, is specified as a bona fide cell cycle-specific substrate of OTUD6B. Stabilization of LIN28B drives MYC expression at G1/S, which in turn allows for rapid S-phase entry. Silencing OTUD6B or LIN28B inhibits multiple myeloma outgrowth in vivo and high OTUD6B expression evolves in patients that progress to symptomatic multiple myeloma and results in an adverse outcome of the disease. Thus, we link proteolytic ubiquitylation with post-transcriptional regulation and nominate OTUD6B as a potential mediator of the MGUS-multiple myeloma transition, a central regulator of MYC, and an actionable vulnerability in multiple myeloma and other tumors with an activated OTUD6B-LIN28B axis.

SUMMARY Objectives The aim of this randomized double-blind controlled clinical trial was to evaluate different protocols for at-home use of 10% hydrogen peroxide in whitening effectiveness and tooth sensitivity. Methods Seventy-two patients were selected according to the inclusion and exclusion criteria, with the upper central incisors having color A2 or darker according to the Vita Classical scale (VITA Zahnfabrik, Bad Säckingen, Germany) and randomized into two groups: 10% hydrogen peroxide applied once daily for 15 minutes (HP 15) or applied once daily for 30 minutes (HP 30). Bleaching was performed for 14 days in both groups. The color was evaluated before bleaching, during bleaching (1st and 2nd weeks), and 1 month after the bleaching treatment using the Vita Classical, Vita Bleachedguide 3D-MASTER, and Vita Easyshade spectrophotometer (VITA Zahnfabrik). Dental sensitivity was recorded by the patients using the numerical rating scale (0–4) and visual analogue scale (0–10 cm). Color data were evaluated by two-way analysis of variance (ANOVA) of repeated measures (group vs. treatment time). The Mann-Whitney test was performed to contrast the means (α=0.05). Tooth sensitivity was assessed by Fisher’s exact test (p=1.00) and intensity of tooth sensitivity was evaluated by the Mann-Whitney test (α=0.05) for both scales. Results A significant whitening effect was observed after 2 weeks of bleaching for all color measurements (p=0.01), with no difference between HP 15 and HP 30 (p>0.05). Also, the absolute risk and intensity of tooth sensitivity were similar (47%; p>0.05). Conclusions The effectiveness and tooth sensitivity of at-home bleaching carried out with 10% hydrogen peroxide applied for 15 minutes or 30 minutes are similar.

ZusammenfassungSelektive PET- oder SPECT- Radiopharmaka sind inzwischen nicht nur für αvβ3, sondern auch weitere der 24 verschiedenen Integrine verfügbar, zum Beispiel α5β1, αvβ6, αvβ8 und α6. Da diese unter anderem auch von verschiedenen Karzinomen und im Zuge von Fibrose exprimiert werden, ist die Vorstellung, dass Integrine nur als Zielstrukturen für die Bildgebung von Angiogenese in Betracht kommen, endgültig überholt. Die derzeit besten Aussichten auf eine breite klinische Anwendung, sowohl diagnostisch als auch therapeutisch, haben derzeit αvβ6-Integrin-Radiopharmaka, da αvβ6 von vielen malignen Krebsarten (v. a. Pankreas-, Plattenepithel-, Basalzell-, Lungen- und Colonkarzinom) überexprimiert wird.

Background. Visual inspection and biopsy is the current standard of care for oral cancer diagnosis, but is subject to misinterpretation and consequently to misdiagnosis. Topically applied PARPi-FL is a molecularly specific, fluorescent contrast-based approach that may fulfil the unmet need for a simple, in vivo, non-invasive, cost-effective, point-of-care method for the early diagnosis of oral cancer. Here, we present results from a phase I safety and feasibility study on fluorescent, topically applied PARPi-FL. Twelve patients with a histologically proven squamous cell carcinoma of the oral cavity (OSCC) gargled a PARPi-FL solution for 60 seconds (15 mL, 100 nM, 250 nM, 500 nM, or 1000 nM), followed by gargling a clearing solution for 60 seconds. Fluorescence measurements of the lesion and surrounding oral mucosa were taken before PARPi-FL application, after PARPi-FL application and after clearing. Blood pressure, oxygen levels, clinical chemistry and CBC were obtained before and after tracer administration. Results. PARPi-FL was well-tolerated by all patients without any safety concerns. When analyzing the fluorescence signal, all malignant lesions showed a significant differential in contrast after administration of PARPi-FL, with the highest increase occurring at the highest dose level (1000 nM), where all patients had a tumor-to-margin fluorescence signal ratio of > 3. A clearing step was essential to increase signal specificity, as it clears unbound PARPi-FL trapped in normal anatomical structures. PARPi-FL tumor cell specificity was confirmed by ex vivo tabletop confocal microscopy. We have demonstrated that the fluorescence signal arose from the nuclei of tumor cells, endorsing our macroscopic findings.Conclusions. A PARPi-FL swish & spit solution is a rapid and non-invasive diagnostic tool that preferentially localizes fluorescent contrast to OSCC. This technique holds promise for the early detection of OSCC based on in vivo optical evaluation and targeted biopsy of suspicious lesions in the oral cavity. Clinicaltrials.gov - NCT03085147, registered on March 21st, 2017.

Synthesis of mixed‐substituted tetraphosphetanes (RP−PtBu)2 (R=Ph (4), Py (5); Py=2‐pyridyl) is achieved from the condensation of dipyrazolylphosphanes RPpyr2 (R=Py (1), Ph (3); pyr=3,5‐dimethylpyrazolyl) as P1‐building block (R−P) and tBuPH2 in an equimolar ratio. Compound 5 is of special interest since the presence of two pyridyl‐substituents as well as the P4‐core allows for a rich coordination chemistry with coinage metal salts [Cu(MeCN)4][OTf], Ag[OTf] and in situ formed [Au(tht)][OTf] (tht=tetrahydrothiophene). Both tetraphosphetanes undergo alkylation reaction with MeOTf to give a series of tetraphosphetanium and tetraphosphetanediium triflate salts with additional methylation of the pyridyl‐moiety in case of 5 resulting in interesting novel cyclic trications. Harsh reaction condition and an excess of MeOTf converts 5 into the cyclic trication [‐P(MePy)PMe2P(MePy)PtBu‐]3+ (13 3+; MePy=1‐methylpyridiniumyl) through the elimination of isobutene. This salt undergoes a complicated rearrangement reaction involving a P−P/P−P bond metathesis to form trication [‐P(MePy)3PtBu‐]3+ (17 3+) when reacted with Me2PPMe2., Choose your donor: A convenient route towards mixed‐substituted tetraphosphetanes was established, enabling the synthesis of (PhP‐PtBu)2 and (PyP‐PtBu)2 (Py=2‐pyridyl) in good yields. The coordination chemistry of (PyP‐PtBu)2 was investigated by reacting it with coinage metal triflate salts. Further methylation studies on both tetraphosphetanes revealed an interesting chemistry of multiply charged, cylic polyphosphanes.

Purpose: Glioblastoma multiforme is a highly aggressive form of brain cancer whose location, tendency to infiltrate healthy surrounding tissue, and heterogeneity significantly limit survival, with scant progress having been made in recent decades. Experimental Design: 123I-MAPi (Iodine-123 Meitner-Auger PARP1 inhibitor) is a precise therapeutic tool composed of a PARP1 inhibitor radiolabeled with an Auger- and gamma-emitting iodine isotope. Here, the PARP inhibitor, which binds to the DNA repair enzyme PARP1, specifically targets cancer cells, sparing healthy tissue, and carries a radioactive payload within reach of the cancer cells' DNA. Results: The high relative biological efficacy of Auger electrons within their short range of action is leveraged to inflict DNA damage and cell death with high precision. The gamma ray emission of 123I-MAPi allows for the imaging of tumor progression and therapy response, and for patient dosimetry calculation. Here we demonstrated the efficacy and specificity of this small-molecule radiotheranostic in a complex preclinical model. In vitro and in vivo studies demonstrate high tumor uptake and a prolonged survival in mice treated with 123I-MAPi when compared with vehicle controls. Different methods of drug delivery were investigated to develop this technology for clinical applications, including convection enhanced delivery and intrathecal injection. Conclusions: Taken together, these results represent the first full characterization of an Auger-emitting PARP inhibitor which demonstrate a survival benefit in mouse models of GBM and confirm the high potential of 123I-MAPi for clinical translation.

For oral, oropharyngeal and oesophageal cancer, the early detection of tumours and of residual tumour after surgery are prognostic factors of recurrence rates and patient survival. Here, we report the validation, in animal models and a human, of the use of a previously described fluorescently labelled small-molecule inhibitor of the DNA-repair enzyme poly(ADP-ribose) polymerase 1 (PARP1) for the detection of cancers of the oral cavity, pharynx and oesophagus. We show that the fluorescent contrast agent can be used to quantify the expression levels of PARP1, and to detect oral, oropharyngeal and oesophageal tumours in mice, pigs and fresh human biospecimens when delivered topically or intravenously. The fluorescent PARP1 inhibitor can also detect oral carcinoma in a patient when applied as a mouthwash, and discriminate, between fresh biopsied samples of the oral tumour and the surgical resection margin with larger than 95% sensitivity and specificity. The PARP1 inhibitor could serve as the basis of a fast and sensitive assay for the early detection, and for the surgical-margin assessment, of epithelial cancers of the upper intestinal tract.

Mutant KRAS is present in over 90% of pancreatic as well as 30-40% of lung and colorectal cancers and is one of the most common oncogenic drivers. Despite decades of research and the recent emergence of isoform-specific KRASG12C-inhibitors, most mutant KRAS isoforms, including the ones frequently associated with pancreatic ductal adenocarcinoma (PDAC), cannot be targeted directly. Moreover, targeting single RAS downstream effectors induces adaptive mechanisms leading to tumor recurrence or resistance. We report here on the combined inhibition of SHP2, a non-receptor tyrosine phosphatase upstream of KRAS, and ERK, a serine/threonine kinase and a key molecule downstream of KRAS in PDAC. This combination shows synergistic anticancer activity in vitro, superior disruption of the MAPK pathway, and significantly increased apoptosis induction compared to single-agent treatments. In vivo, we demonstrate good tolerability and efficacy of the combination. Concurrent inhibition of SHP2 and ERK induces significant tumor regression in multiple PDAC mouse models. Finally, we show evidence that 18F-FDG PET scans can be used to detect and predict early drug responses in animal models. Based on these compelling results, we will investigate this drug combination in a clinical trial (SHERPA, SHP2 and ERK inhibition in pancreatic cancer, NCT04916236), enrolling patients with KRAS-mutant PDAC.

High levels of reactive oxygen species (ROS) in tumors have been shown to exert anti-tumor activity, leading to the concept of ROS induction as therapeutic strategy. The organometallic compound ferrocene (Fc) generates ROS through a reversible one-electron oxidation. Incorporation of Fc into a tumor-targeting, bioactive molecule can enhance its therapeutic activity and enable tumor specific delivery. Therefore, we conjugated Fc to five synthetic, Arg-Gly-Asp (RGD)-based integrin binding ligands to enable targeting of the cell adhesion and signaling receptor integrin subtypes αvβ3, α5β1, or αvβ6, which are overexpressed in various, distinct tumors. We designed and synthesized a library of integrin-ligand-ferrocene (ILF) derivatives and showed that ILF conjugates maintained the high integrin affinity and selectivity of their parent ligands. A thorough biological characterization allowed us to identify the two most promising ligands, an αvβ3 (L2b) and an αvβ6 (L3b) targeting ILF, which displayed selective integrin-dependent cell uptake and pronounced ferrocene-mediated anti-tumor effects in vitro, along with increased ROS production and DNA damage. Hence, ILFs are promising candidates for the selective, tumor-targeted delivery of ferrocene to maximize its anti-cancer efficacy and minimize systemic toxicity, thereby improving the therapeutic window of ferrocene compared to currently used non-selective anti-cancer drugs.