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

A portable microfluidic electrochemical sensor with nonlinear fit strategy for wide-range uric acid detection

In this work, we have developed a portable electrochemical sensing device controlled by a smartphone through Bluetooth, which was applied for regular uric acid (UA) monitoring in prevention and healthcare management. On the core lies a nonlinear fitting model (I = k·(N − exp(−K·cbulk + b))) which is proposed for the first time and given from the differential equation based on the theoretical analysis. We believe this model can reflect the intrinsic relationship between the obtained current and the bulk analyte concentration, therefore leading to an expanding UA detection range (10-fold) compared to the widely used linear standard curve model and reducing the requirements for the modification of the electrode materials. Besides, the integration of the microfluidic chip promoted the removal of UA oxidation products at the electrode surface, yielding excellent sensing stability with a relative standard deviation (RSD) lower than 1 % during 10 consecutive runs. Consequently, we acquired the relationship between Square Wave Voltammetry (SWV) current and UA concentration (I = 15.27*(2.05 − exp(−0.00108*(cbulk − 663.5))), R2 = 0.9999) in the range of 5 to 1000 μM with a limit of detection (LoD) of 2.4 μM using a completely unmodified screen-printed carbon electrode (SPCE). Proof of concept experiments using 25× diluted human urine spiked with UA yielded a recovery rate of 87.5–101.4 % and a satisfactory selectivity result, which is within the value expected for clinical use, indicating the potential of the developed instrument and nonlinear fitting model for urinary UA detection.</p

TAO Conceptual Design Report: A Precision Measurement of the Reactor Antineutrino Spectrum with Sub-percent Energy Resolution

The Taishan Antineutrino Observatory (TAO, also known as JUNO-TAO) is a satellite experiment of the Jiangmen Underground Neutrino Observatory (JUNO). A ton-level liquid scintillator detector will be placed at about 30 m from a core of the Taishan Nuclear Power Plant. The reactor antineutrino spectrum will be measured with sub-percent energy resolution, to provide a reference spectrum for future reactor neutrino experiments, and to provide a benchmark measurement to test nuclear databases. A spherical acrylic vessel containing 2.8 ton gadolinium-doped liquid scintillator will be viewed by 10 m^2 Silicon Photomultipliers (SiPMs) of &gt;50% photon detection efficiency with almost full coverage. The photoelectron yield is about 4500 per MeV, an order higher than any existing large-scale liquid scintillator detectors. The detector operates at -50 degree C to lower the dark noise of SiPMs to an acceptable level. The detector will measure about 2000 reactor antineutrinos per day, and is designed to be well shielded from cosmogenic backgrounds and ambient radioactivities to have about 10% background-to-signal ratio. The experiment is expected to start operation in 2022

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

Microfluidic devices for the isolation and label-free identification of circulating tumor cells

The isolation and identification of circulating tumor cells (CTCs) from blood samples provides a great opportunity for prevention of metastatic cancer and assist early diagnosis and treatment of cancer. However, separation and recovery of CTCs using conventional analytical technologies remains challenging due to the low concentration of CTCs in blood. Microfluidics technologies have recently been proposed as advanced platform for isolation of CTCs owing to its ease of integration, rapid and continuous processing, and separation of cells along multiple geometric possibilities in manipulating micron-sized particles. This review provides a critical and comprehensive outlook to recent advancements, challenges, and opportunities of microfluidic, label-free strategies for highly CTCs isolation from biological samples. This includes a detailed summary of microfluidic-based methods for cell sorting, which have been organized into active and passive methods with a detailed discussion on the practical applications of each process in improving the performance for isolation of CTCs, looking at their working principles and respective advantages. We elaborated on some of these methods’ current limitations and prospective opportunities, which we believe will help to drive further innovations and adoption of microfluidic-based CTCs isolation, with a large potential for clinical applications in cancer treatment and diagnostics and fundamental study of metastatic processes.</p

ORY-1001 Suppresses Cell Growth and Induces Apoptosis in Lung Cancer Through Triggering HK2 Mediated Warburg Effect

ORY-1001, an inhibitor of covalent lysine (K)-specific demethylase 1A (KDM1A), has been used as a therapy for the treatment of acute leukemia. However, the underlying mechanisms of anticancer are still not fully elucidated. Here, we report that KDM1A is highly expressed in lung cancers, where it appears to drive aggressive growth. Furthermore, lung cancer patients with higher KDM1A levels have worse survival outcomes than patients with lower KDM1A levels. Interestingly, ORY-1001significantly inhibited the cell proliferation, colony formation, cell cycle, and induced apoptosis, by regulating the Warburg effect through controlling Hexokinases 2 (HK2) expression. In summary, these results indicate that ORY-1001 could inhibit the growth of lung cancer cells via regulating the Warburg effect by controlling HK2