Design, Synthesis, and Biological Evaluation of Potent and Selective PROTAC Degraders of Oncogenic KRASG12D.

KRASG12D, the most frequent KRAS oncogenic mutation, is a promising target for cancer therapy. Herein, we report the design, synthesis, and biological evaluation of a series of KRASG12D PROTACs by connecting the analogues of MRTX1133 and the VHL ligand. Structural modifications of the linker moiety and KRAS inhibitor part suggested a critical role of membrane permeability in the degradation activity of the KRASG12D PROTACs. Mechanism studies with the representative compound 8o demonstrated that the potent, rapid, and selective degradation of KRASG12D induced by 8o was via a VHL- and proteasome-dependent manner. This compound selectively and potently suppressed the growth of multiple KRASG12D mutant cancer cells, displayed favorable pharmacokinetic and pharmacodynamic properties in mice, and showed significant antitumor efficacy in the AsPC-1 xenograft mouse model. Further optimization of 8o appears to be promising for the development of a new chemotherapy for KRASG12D-driven cancers as the complementary therapeutic strategy to KRAS inhibition.

Discovery of a Potent, Cooperative, and Selective SOS1 PROTAC ZZ151 with In Vivo Antitumor Efficacy in KRAS-Mutant Cancers.

The linker moiety of a proteolysis-targeting chimera (PROTAC) molecule plays a critical role in modulating the degradation activity, target selectivity, and physico-chemical properties. However, the basics and underlying mechanisms of chemical modifications of the linker structure causing dramatic changes in the PROTAC degradation activity warrant further investigation. Herein, we report the design and characterization of a highly potent and selective SOS1 PROTAC ZZ151. After systematically modifying the linker length and composition, we observed that subtle modification of just one atom of the linker moiety of ZZ151 resulted in remarkable changes in the formation of the ternary complex and thus dramatically affected the degradation activities. ZZ151 quickly, specifically, and effectively induced SOS1 degradation; displayed potent antiproliferation activities against a broad panel of KRAS mutant-driven cancer cells; and showed superior anticancer activities in the KRASG12D- and G12V-mutant xenografts in mice. ZZ151 is a promising lead for developing new chemotherapies targeting KRAS mutants.

Design, Synthesis, and Bioevaluation of Pyrido[2,3-d]pyrimidin-7-ones as Potent SOS1 Inhibitors.

The use of small molecular modulators to target the guanine nucleotide exchange factor SOS1 has been demonstrated to be a promising strategy for the treatment of various KRAS-driven cancers. In the present study, we designed and synthesized a series of new SOS1 inhibitors with the pyrido[2,3-d]pyrimidin-7-one scaffold. One representative compound 8u showed comparable activities to the reported SOS1 inhibitor BI-3406 in both the biochemical assay and the 3-D cell growth inhibition assay. Compound 8u obtained good cellular activities against a panel of KRAS G12-mutated cancer cell lines and inhibited downstream ERK and AKT activation in MIA PaCa-2 and AsPC-1 cells. In addition, it displayed synergistic antiproliferative effects when used in combination with KRAS G12C or G12D inhibitors. Further modifications of the new compounds may give us a promising SOS1 inhibitor with favorable druglike properties for use in the treatment of KRAS-mutated patients.

The spectral background problem of portable fiber Raman instruments and a solution for the on-site detection of extremely weak signals.

High sensitivity and low spectral background of the Raman instruments are essential for the analysis of weak Raman-active samples. The design of the present portable fiber Raman probes leads to an interferential background that limits the application of the portable Raman technique in many situations. In this work, the spectral backgrounds of commercial probes with different optical configurations were analyzed, and the results indicated the backgrounds arose from the objective lenses and the dichroic filters. A universal solution was proposed, and a flat and clean background was achieved by utilizing the longpass optical configuration and an off-axis parabolic mirror. Combined with a high-throughput spectrometer with a research-grade CCD, this portable Raman equipment was successfully applied to identify the corrosion products on the surface of uranium (with extremely weak Raman signals). The developed portable Raman instrument with high sensitivity and low background is anticipated to be very useful for the on-site Raman detection of weak signals.

A practical wide-field Raman imaging method with high spectral and spatial resolution.

Raman imaging has a great advantage in characterizing inhomogeneous systems. A practical wide-field Raman imaging platform is developed that shows major improvements on imaging speed, sensitivity, and resolution. Different from the traditional Raman imaging systems using the wavelength-fixed lasers and the chromatic dispersion devices, this system adopts an inverted architecture, integrated with a tunable laser and the wavelength-fixed filters. Owing to the high transmission of the fixed filters, the imaging sensitivity can be improved 5-10 times in comparison to the present wide-field Raman imaging setups using liquid-crystal tunable filters. Via combining with the high-power tunable laser, Raman images could be obtained in minutes and the Raman shift of the images could be tuned easily and accurately. The resolution of this system can reach 1.5 cm-1 in the spectrum and 490 nm in space, which could provide more fingerprint details of the analytes. This effective Raman imaging method allowing us to see chemical spatial variations on microscale is anticipated to be widely applied in scientific research fields.

Transcriptome-referenced association study of clove shape traits in garlic.

Genome-wide association studies are a powerful approach for identifying genes related to complex traits in organisms, but are limited by the requirement for a reference genome sequence of the species under study. To circumvent this problem, we propose a transcriptome-referenced association study (TRAS) that utilizes a transcriptome generated by single-molecule long-read sequencing as a reference sequence to score population variation at both transcript sequence and expression levels. Candidate transcripts are identified when both scores are associated with a trait and their potential interactions are ascertained by expression quantitative trait loci analysis. Applying this method to characterize garlic clove shape traits in 102 landraces, we identified 22 candidate transcripts, most of which showed extensive interactions. Eight transcripts were long non-coding RNAs (lncRNAs), and the others were proteins involved mainly in carbohydrate metabolism, protein degradation, etc. TRAS, as an efficient tool for association study independent of a reference genome, extends the applicability of association studies to a broad range of species.

Substantial performance improvement in inverted polymer light-emitting diodes via surface plasmon resonance induced electrode quenching control.

Inverted-type polymer light-emitting diodes with Au nanoparticles modified ITO cathode has exhibited improved brightness from 5900 to 15,000 cd m(-2) (1.5-fold enhancement) and enhanced luminous efficiency from 4.4 to 10.5 cd A(-1) (1.4-fold enhancement), when greenish emissive polymer-P-PPV was applied as active layer. Both the experimental and theoretical results show that it is mainly attributed to effective overlapping between local surface plasmon resonance induced by Au nanopartices and excitons quenching region at ZnO/P-PPV interface, which makes originally electrode-quenched excitons emissive and increases excitons efficiency.

Note: Raman microspectroscopy integrated with fluorescence and dark field imaging.

A Raman detection platform integrated with both fluorescence and dark field microscopes was built for in situ Raman detection with the assistance of fluorescence and dark field imaging to locate the target micro regions. Cells and organelles can be easily found via fluorescence imaging with labeling techniques. Besides, nano-sized particles could be observed and located by dark field microscopes. Therefore, comparing with the commercial Raman spectrometers, much more researches based on Raman spectroscopy could be carried out on this integrated Raman platform, especially in the fields of analyzing biological tissues and subwavelength samples.

Evanescent field excited plasmonic nano-antenna for improving SERS signal.

The purpose of this work is to develop a high-performance surface-enhanced Raman scattering (SERS) substrate with high light harvesting and SERS emitting efficiencies based on the principle of a plasmonic nano-antenna. A prism/Ag nanowell array was designed and constructed based on the Kretschmann configuration. Almost 100% of the incident light can be absorbed at the resonance angle. A strong electromagnetic (EM) field is generated in the near field along the rim and at the center of the nanowells by means of the localized energy of surface plasmons (SPs), resulting in the electric field being enhanced 300 times at the Ag surface. The co-enhancement of localized SPs and propagating SPs was achieved in this substrate. The plasmonic nano-antenna structure can also redirect the SERS, which benefits the SERS collection. The SERS sensitivity on this configuration was improved by about 40 times compared with that on a conventional Kretschmann configuration with a flat Ag film. This SERS substrate design with full consideration on the tuning of the EM field in the near field and SERS space distribution in the far field could give insight into the design of a new generation of SERS substrates.

Note: Mobile micro-Raman analyzer integrated with a lab-on-a-chip.

A mobile micro-Raman microfluidic analyzer was designed and built for label-free, nondestructive, fingerprint detection of samples on microfluidic chip systems. It mainly includes an optical module (including the Raman detection system and the microscopic imaging system), 3-axis stages with step motors and other auxiliary circuits. The setup of the analyzer was designed with fully considering the characters of the microfluidics system. The experimental results prove that this microfluidics analyzer is practical and very convenient to use. This micro-Raman microfluidics analyzer with high performance to cost ratio has wide application potential in lab-on-a-chip fields as a powerful analytical tool.

Overexpression of a GmGBP1 ortholog of soybean enhances the responses to flowering, stem elongation and heat tolerance in transgenic tobaccos.

Soybean is a typical short-day crop, and its photoperiodic and gibberellin (GA) responses for the control of flowering are critical to seed yield. The GmGBP1 mRNA abundance in leaves was dramatically increased in short-days (SDs) compared to that in long-days in which it was consistently low at all time points from 0 to 6 days (days after transfer to SDs). GmGBP1 was highly expressed in leaves and exhibited a circadian rhythm in SDs. Ectopic overexpression of GmGBP1 in tobaccos caused photoperiod-insensitive early flowering by increasing NtCO mRNA levels. GmGBP1 mRNA abundance was also increased by GAs. Transgenic GmGBP1 overexpressing (-ox) tobacco plants exhibited increased GA signaling-related phenotypes including flowering and plant height promotion. Furthermore, the hypocotyl elongation, early-flowering and longer internode phenotypes were largely accelerated by GA3 application in the GmGBP1-ox tobacco seedlings. Being consistent, overexpression of GmGBP1 resulted in significantly enhanced GA signaling (evidenced suppressed expression of NtGA20ox) both with and without GA treatments. GmGBP1 was a positive regulator of both photoperiod and GA-mediated flowering responses. In addition, GmGBP1-ox tobaccos were hypersensitive to ABA, salt and osmotic stresses during seed germination. Heat-inducible GmGBP1 also enhanced thermotolerance in transgenic GmGBP1-ox tobaccos during seed germination and growth. GmGBP1 protein was localized in the nucleus. Analyses of a series of 5'-deletions of the GmGBP1 promoter suggested that several cis-acting elements, including P-BOX, TCA-motif and three HSE elements necessary to induce gene expression by GA, salicic acid and heat stress, were specifically localized in the GmGBP1 promoter region.

Hierarchical structural nanopore arrays fabricated by pre-patterning aluminum using nanosphere lithography.

A highly ordered and hierarchical structural nanopore array is fabricated via anodizing a pre-patterned aluminum foil under an optimized voltage. A pre-patterned hexagonal nanoindentation array on an aluminum substrate is prepared via the nanosphere lithography method. This pattern leads to an elaborate nanochannel structure with seven nanopores in each nanoindentation after anodization treatment. The structure achieved in our study is new, interesting, and likely to be applied in photonic devices.