Activation of the p53 Transcriptional Program Sensitizes Cancer Cells to Cdk7 Inhibitors

Cdk7, the CDK-activating kinase and transcription factor IIH component, is a target of inhibitors that kill cancer cells by exploiting tumor-specific transcriptional dependencies. However, whereas selective inhibition of analog-sensitive (AS) Cdk7 in colon cancer-derived cells arrests division and disrupts transcription, it does not by itself trigger apoptosis efficiently. Here, we show that p53 activation by 5-fluorouracil or nutlin-3 synergizes with a reversible Cdk7asinhibitor to induce cell death. Synthetic lethality was recapitulated with covalent inhibitors of wild-type Cdk7, THZ1, or the more selective YKL-1-116. The effects were allele specific; a CDK7asmutation conferred both sensitivity to bulky adenine analogs and resistance to covalent inhibitors. Non-transformed colon epithelial cells were resistant to these combinations, as were cancer-derived cells with p53-inactivating mutations. Apoptosis was dependent on death receptor DR5, a p53 transcriptional target whose expression was refractory to Cdk7 inhibition. Therefore, p53 activation induces transcriptional dependency to sensitize cancer cells to Cdk7 inhibition. Kalan et al. find that activation of the p53 tumor suppressor protein in human colon cancer-derived cells can induce transcriptional dependency on Cdk7, analogous to constitutive dependencies described in other tumors driven by oncogenic transcription factors. This work provides a proof of concept for combining p53-activating agents with Cdk7 inhibitors to elicit synthetic lethality. Keywords: Cdk7; p53; colon cancer; synthetic lethality; transcription; 5-fluorouracil; nutlin-3; apoptosis; chemical genetics; CDK inhibitorNational Institutes of Health (U.S.) (Grant HG002668

Covalent targeting of remote cysteine residues to develop CDK12 and CDK13 inhibitors

Cyclin-dependent kinases 12 and 13 (CDK12 and CDK13) play critical roles in the regulation of gene transcription. However, the absence of CDK12 and CDK13 inhibitors has hindered the ability to investigate the consequences of their inhibition in healthy cells and cancer cells. Here we describe the rational design of a first-in-class CDK12 and CDK13 covalent inhibitor, THZ531. Co-crystallization of THZ531 with CDK12–cyclin K indicates that THZ531 irreversibly targets a cysteine located outside the kinase domain. THZ531 causes a loss of gene expression with concurrent loss of elongating and hyperphosphorylated RNA polymerase II. In particular, THZ531 substantially decreases the expression of DNA damage response genes and key super-enhancer-associated transcription factor genes. Coincident with transcriptional perturbation, THZ531 dramatically induced apoptotic cell death. Small molecules capable of specifically targeting CDK12 and CDK13 may thus help identify cancer subtypes that are particularly dependent on their kinase activities.United States. National Institutes of Health (HG002668)United States. National Institutes of Health (CA109901

Covalent targeting of remote cysteine residues to develop CDK12 and CDK13 inhibitors

Cyclin-dependent kinases 12 and 13 (CDK12 and CDK13) play critical roles in the regulation of gene transcription. However, the absence of CDK12 and CDK13 inhibitors has hindered the ability to investigate the consequences of their inhibition in healthy cells and cancer cells. Here we describe the rational design of a first-in-class CDK12 and CDK13 covalent inhibitor, THZ531. Co-crystallization of THZ531 with CDK12–cyclin K indicates that THZ531 irreversibly targets a cysteine located outside the kinase domain. THZ531 causes a loss of gene expression with concurrent loss of elongating and hyperphosphorylated RNA polymerase II. In particular, THZ531 substantially decreases the expression of DNA damage response genes and key super-enhancer-associated transcription factor genes. Coincident with transcriptional perturbation, THZ531 dramatically induced apoptotic cell death. Small molecules capable of specifically targeting CDK12 and CDK13 may thus help identify cancer subtypes that are particularly dependent on their kinase activities.United States. National Institutes of Health (HG002668)United States. National Institutes of Health (CA109901

Real-time Monitoring for the Next Core-Collapse Supernova in JUNO

Core-collapse supernova (CCSN) is one of the most energetic astrophysical events in the Universe. The early and prompt detection of neutrinos before (pre-SN) and during the SN burst is a unique opportunity to realize the multi-messenger observation of the CCSN events. In this work, we describe the monitoring concept and present the sensitivity of the system to the pre-SN and SN neutrinos at the Jiangmen Underground Neutrino Observatory (JUNO), which is a 20 kton liquid scintillator detector under construction in South China. The real-time monitoring system is designed with both the prompt monitors on the electronic board and online monitors at the data acquisition stage, in order to ensure both the alert speed and alert coverage of progenitor stars. By assuming a false alert rate of 1 per year, this monitoring system can be sensitive to the pre-SN neutrinos up to the distance of about 1.6 (0.9) kpc and SN neutrinos up to about 370 (360) kpc for a progenitor mass of 30$M_{\odot}$ for the case of normal (inverted) mass ordering. The pointing ability of the CCSN is evaluated by using the accumulated event anisotropy of the inverse beta decay interactions from pre-SN or SN neutrinos, which, along with the early alert, can play important roles for the followup multi-messenger observations of the next Galactic or nearby extragalactic CCSN.Comment: 24 pages, 9 figure

Potential of Core-Collapse Supernova Neutrino Detection at JUNO

JUNO is an underground neutrino observatory under construction in Jiangmen, China. It uses 20kton liquid scintillator as target, which enables it to detect supernova burst neutrinos of a large statistics for the next galactic core-collapse supernova (CCSN) and also pre-supernova neutrinos from the nearby CCSN progenitors. All flavors of supernova burst neutrinos can be detected by JUNO via several interaction channels, including inverse beta decay, elastic scattering on electron and proton, interactions on C12 nuclei, etc. This retains the possibility for JUNO to reconstruct the energy spectra of supernova burst neutrinos of all flavors. The real time monitoring systems based on FPGA and DAQ are under development in JUNO, which allow prompt alert and trigger-less data acquisition of CCSN events. The alert performances of both monitoring systems have been thoroughly studied using simulations. Moreover, once a CCSN is tagged, the system can give fast characterizations, such as directionality and light curve

Detection of the Diffuse Supernova Neutrino Background with JUNO

As an underground multi-purpose neutrino detector with 20 kton liquid scintillator, Jiangmen Underground Neutrino Observatory (JUNO) is competitive with and complementary to the water-Cherenkov detectors on the search for the diffuse supernova neutrino background (DSNB). Typical supernova models predict 2-4 events per year within the optimal observation window in the JUNO detector. The dominant background is from the neutral-current (NC) interaction of atmospheric neutrinos with 12C nuclei, which surpasses the DSNB by more than one order of magnitude. We evaluated the systematic uncertainty of NC background from the spread of a variety of data-driven models and further developed a method to determine NC background within 15\% with {\it{in}} {\it{situ}} measurements after ten years of running. Besides, the NC-like backgrounds can be effectively suppressed by the intrinsic pulse-shape discrimination (PSD) capabilities of liquid scintillators. In this talk, I will present in detail the improvements on NC background uncertainty evaluation, PSD discriminator development, and finally, the potential of DSNB sensitivity in JUNO

Synthesis and Methodology Development Involving Strained Heterocycles and Preparation of Immunodulatory Glycosphingolipids

Strained heterocycles serve as important synthetic intermediates in organic chemistry, as well as motifs for natural products. In this thesis, we report a short, straightforward synthesis of the first psico-oxetanocin analogue of the powerful antiviral natural product, oxetanocin A, from 2-methyleneoxetanes. This unusual class of strained heterocycles was derived from β-lactones prepared by the carbonylation of epoxides. F+- mediatednucleobase incorporation provided the corresponding nucleosides in good yield but with low diastereoselectivity. The exploitation of anchimeric assistance to increase the selectivity was attempted but not fruitful. A range of 2-methyleneoxetane and related 2-methylenetetrahydrofuran substrates was prepared to explore the basis for this. With one exception, these substrates also showed little stereoselectivity in nucleobase incorporation. Computational studies were undertaken to examine if neighboring group participation involving fused [4.2.0] or [4.3.0] intermediates is favorable. ^ We have also studied the olefin cross metathesis reactions involving another class of strained heterocycles, a-methylene-13-lactams. This methodology showed good tolerance of a variety of functionalities, providing α-alkylidene-β-lactams in good to excellent yields with either Grubbs or Hoveyda-Grubbs 2 nd catalysts. Electron-poor α-methylene-β-lactams undergo cross-metathesis more rapidly and efficiently than more electron-rich analogues. Significantly, tetrasubstituted alkenes have for the first time been accessed by CM reactions. ^ Glycolipids, particularly α-galactosylceramides (α-GalCers), can stimulate NKT cells, which play a central role in regulating immune responses. Tremendous efforts have been devoted to the understanding of the activating process and the fine tuning of the cytokine release (Thl vs. Th2) by altering either the sugar or lipid portions of KRN7000, a principle α-GalCer for the study of NKT cell stimulation. One theory suggested that glycolipids which elicit high Thl bias require antigen presenting cells (APCs), which were shown to be essential to truncate (α1-2) digalactosylceramides to KRN7000. We synthesized the direct disaccharide analogue of a Thl-biasing α-GalCer, hypothesizing that the Thl bias may be enhanced by adding a monosaccharide in a position known to be subject to glycosidase cleavage. The armed-disarmed concept in sugar chemistry has been used for the key glycosylation steps to achieve better reactivity and high α-selectivity.

Multi-Dimensional Interval Number Decision Model Based on Mahalanobis-Taguchi System with Grey Entropy Method and Its Application in Reservoir Operation Scheme Selection

In decision-making with interval numbers, there are problems such as how to reduce the loss of decision information to improve decision accuracy and the difficulty of using interval numbers for sorting. On the basis of fully considering the subjective and objective weights of indexes, the grey entropy method (GEM) is improved by taking advantage of the Mahalanobis-Taguchi System (MTS) in which the orthogonal design has few tests but much obtained information, and the Mahalanobis distance can reflect the correlation between indexes. Then, the signal-to-noise ratio is integrated with the improved degree of balance and approach, and a multi-dimensional interval number decision model based on MTS and GEM is put forth. This model is applied to selecting the optimal scheme of controlling the Pankou reservoir’s water level in flood season. Compared with the decision results of other methods, the optimal scheme selected by the proposed model can achieve greater benefits within an acceptable risk range and thus better coordinate the balance between risk and benefit, which verifies the feasibility and validity of the model

Robust Reconstruction of Electrocardiogram Using Photoplethysmography: A Subject-Based Model

Electrocardiography and photoplethysmography are non-invasive techniques that measure signals from the cardiovascular system. While the cycles of the two measurements are highly correlated, the correlation between the waveforms has rarely been studied. Measuring the photoplethysmogram (PPG) is much easier and more convenient than the electrocardiogram (ECG). Recent research has shown that PPG can be used to reconstruct the ECG, indicating that practitioners can gain a deep understanding of the patients' cardiovascular health using two physiological signals (PPG and ECG) while measuring only PPG. This study proposes a subject-based deep learning model that reconstructs an ECG using a PPG and is based on the bidirectional long short-term memory model. Because the ECG waveform may vary from subject to subject, this model is subject-specific. The model was tested using 100 records from the MIMIC III database. Of these records, 50 had a circulatory disease. The results show that a long ECG signal could be effectively reconstructed from PPG, which is, to our knowledge, the first attempt in this field. A length of 228 s of ECG was constructed by the model, which was trained and validated using 60 s of PPG and ECG signals. To segment the data, a different approach that segments the data into short time segments of equal length (and that do not rely on beats and beat detection) was investigated. Segmenting the PPG and ECG time series data into equal segments of 1-min width gave the optimal results. This resulted in a high Pearson's correlation coefficient between the reconstructed 228 s of ECG and referenced ECG of 0.818, while the root mean square error was only 0.083 mV, and the dynamic time warping distance was 2.12 mV per second on average.ISSN:1664-042