Impaired cell fate through gain-of-function mutations in a chromatin reader.

Modifications of histone proteins have essential roles in normal development and human disease. Recognition of modified histones by 'reader' proteins is a key mechanism that mediates the function of histone modifications, but how the dysregulation of these readers might contribute to disease remains poorly understood. We previously identified the ENL protein as a reader of histone acetylation via its YEATS domain, linking it to the expression of cancer-driving genes in acute leukaemia1. Recurrent hotspot mutations have been found in the ENL YEATS domain in Wilms tumour2,3, the most common type of paediatric kidney cancer. Here we show, using human and mouse cells, that these mutations impair cell-fate regulation by conferring gain-of-function in chromatin recruitment and transcriptional control. ENL mutants induce gene-expression changes that favour a premalignant cell fate, and, in an assay for nephrogenesis using murine cells, result in undifferentiated structures resembling those observed in human Wilms tumour. Mechanistically, although bound to largely similar genomic loci as the wild-type protein, ENL mutants exhibit increased occupancy at a subset of targets, leading to a marked increase in the recruitment and activity of transcription elongation machinery that enforces active transcription from target loci. Furthermore, ectopically expressed ENL mutants exhibit greater self-association and form discrete and dynamic nuclear puncta that are characteristic of biomolecular hubs consisting of local high concentrations of regulatory factors. Such mutation-driven ENL self-association is functionally linked to enhanced chromatin occupancy and gene activation. Collectively, our findings show that hotspot mutations in a chromatin-reader domain drive self-reinforced recruitment, derailing normal cell-fate control during development and leading to an oncogenic outcome.

High-power 100 W Kerr-lens mode-locked ring-cavity femtosecond Yb:YAG thin-disk oscillator.

High-power femtosecond pulses delivered at a high-repetition rate will aid machining throughput and improve signal-to-noise ratios for sensitive measurements. Here we demonstrate a Kerr-lens mode-locked femtosecond Yb:YAG ring-cavity thin-disk oscillator with a multi-pass scheme for the laser beam. With four passes through the thin disk, 175-fs pulses were delivered from the oscillator at an average power of 71.5 W and a repetition rate of 65.3 MHz. The corresponding intra-cavity peak power of 110 MW is ample for intra-cavity nonlinear conversion into more exotic wavelength ranges. With six passes, the average output power reached 101.3 W. To the best of our knowledge, this is the highest average output power of any mode-locked ring laser. These results confirm the viability of using multi-pass configuration on a thin-disk ring oscillator for high-throughput femtosecond applications.

205-240 GHz free-space-to-fiber mode adapter with an 80% mode conversion efficiency.

In this work, we propose an integrated terahertz mode adapter that couples broadband terahertz radiation from free-space to hollow-core fiber with a high mode conversion efficiency (Gaussian beam-to-T E 01) of up to 80%. The adapter consists of a pyramidal horn antenna, a broadband mode converter, and a conical horn. The simulation results indicate that the T E 01 mode in the hollow-core fiber can be efficiently excited by the terahertz mode adapter. The terahertz mode adapter successfully achieved a wide operating bandwidth of more than 15.7% ranging from 205 to 240 GHz in our simulation.

Sub-100-fs Kerr-lens mode-locked Yb:YAG ring-cavity thin-disk oscillator.

Ultrafast ring-cavity thin-disk oscillators combine high output power with the flexibility of generating output either unidirectionally or bidirectionally. Here, we report a Kerr-lens mode-locked ring-cavity Yb:YAG thin-disk oscillator delivering unidirectional 89-fs pulses by inducing additional spectral broadening with nonlinear plates. This is the shortest pulse duration for a ring-cavity mode-locked thin-disk oscillator. Bidirectional mode-locking was also realized. These results lay the foundation for the more efficient generation of high-order harmonics at MHz repetition rates and high-power dual frequency combs.

60.4 W continuous-wave single-frequency laser output from a two-stage Nd:YAG single-crystal fiber amplifier.

A Nd:YAG single-crystal fiber amplifier for the amplification of continuous-wave single-frequency laser end-pumped by a laser diode (LD) is investigated. With a two-stage amplification configuration, an output power of 60.4 W under the total incident pump power of 200 W is achieved, which is, to our knowledge, the highest power from a continuous-wave single-frequency laser achieved with a single-crystal fiber scheme. The extraction efficiency reaches 41.6% in the second amplification stage, which is comparable with Innoslab amplifiers. The beam quality factors M2 at the maximum output power in the horizontal and vertical direction are measured to be 1.51 and 1.38, respectively. The long-term power instability for 1 hour is 0.97%.

Sub-60-fs, compact 1100-nm fiber laser system based on double-pass pre-chirp managed amplification.

An ytterbium-doped, single-stage, double-pass nonlinear fiber amplification system was fabricated for amplifying an 1100-nm mode-locking fiber laser. Pre-chirp managed amplification (PCMA) was applied in realizing the nonlinear amplification process with an all-polarization-maintaining (PM) fiber construction. The system can deliver 19.8-nJ, 58.7-fs, 24.4-MHz amplified signal pulses with a 10-dB spectral range spanning from 1049 nm to 1130 nm. Further experimental investigations were conducted in exploring the dynamics of the double-pass nonlinear amplification process. This compact 1100-nm ultrafast fiber laser can be implemented for multi-photon microscopy (MPM) with deep penetration depth.

Repression of human and mouse brain inflammaging transcriptome by broad gene-body histone hyperacetylation.

Brain "inflammaging," a low-grade and chronic inflammation, is a major hallmark for aging-related neurodegenerative diseases. Here, by profiling H3K27ac and gene expression patterns in human and mouse brains, we found that age-related up-regulated (Age-Up) and down-regulated (Age-Down) genes have distinct H3K27ac patterns. Although both groups show promoter H3K27ac, the Age-Up genes, enriched for inflammation-related functions, are additionally marked by broad H3K27ac distribution over their gene bodies, which is progressively reduced during aging. Age-related gene expression changes can be predicted by gene-body H3K27ac level. Contrary to the presumed transcription activation function of promoter H3K27ac, we found that broad gene-body hyper H3K27ac suppresses overexpression of inflammaging genes. Altogether, our findings revealed opposite regulations by H3K27ac of Age-Up and Age-Down genes and a mode of broad gene-body H3K27ac in repressing transcription.

Long-term stable supercontinuum generation and watt-level transmission in liquid-core optical fibers.

Due to their unique properties such as transparency, tunability, nonlinearity, and dispersion flexibility, liquid-core fibers represent an important approach for future coherent mid-infrared light sources. However, the damage thresholds of these fibers are largely unexplored. Here we report on the generation of soliton-based supercontinua in carbon disulfide (CS2) liquid-core fibers at average power levels as high as 0.5 W operating stably for a long term (>70 h) without any kind of degradation or damage. Additionally, we also show stable high-power pulse transmission through liquid-core fibers exceeding 1 W of output average power for both CS2 and tetrachloroethylene as core materials.

Watt-level 193 nm source generation based on compact collinear cascaded sum frequency mixing configuration.

A compact, dual-stage, collinear-cascaded sum-frequency mixing configuration is presented for generating 193 nm sources. Due to the less-introduced, deep-ultraviolet optical components, the system is less prone to damage. In our proof-of-concept experiments, a 1030 nm laser and a 1553 nm laser synchronized to each other were used as drivers and an average power of ~0.7 W was obtained. For comparison, the noncollinear configuration gave an average power of 0.77 W. The difference of 0.07 W is attributed to the spatial walk-off inside the cesium lithium borate (CLBO) crystal, confirmed by indirect visualization. A possible way to overcome the small gap of 0.07 W is proposed for future work.

High-power and high-conversion efficiency deep ultraviolet (DUV) laser at 258 nm generation in the CsLiB6O10 (CLBO) crystal with a beam quality of M2<1.5.

We report a 10 kHz, 10.5 W deep ultraviolet (DUV) laser at 258 nm with a pulse duration of 3 ns by fourth-harmonic generation (FHG) in a CsLiB6O10 crystal with high-conversion efficiency. The fundamental laser is at 1030 nm with 35 W output power consisting of a Yb:YAG ceramics thin rod amplifier. The M2 of the DUV laser was confirmed to be <1.5. To the best of our knowledge, this is the best beam quality for DUV laser power higher than 10 W generated by FHG of a solid-state laser, as well as the highest conversion efficiency of 31% from infrared to DUV so far.

300-mW narrow-linewidth deep-ultraviolet light generation at 193 nm by frequency mixing between Yb-hybrid and Er-fiber lasers.

A narrow-linewidth, high average power deep-ultraviolet (DUV) coherent laser emitting at 193 nm is demonstrated by frequency mixing a Yb-hybrid laser with an Er-fiber laser. The Yb-hybrid laser consists of Yb-fiber lasers and an Yb:YAG amplifier. The average output power of the 193 nm laser is 310 mW at 6 kHz, which corresponds to a pulse energy of 51 µJ. To the best of our knowledge, this is the highest average power and pulse energy ever reported for a narrow-linewidth 193 nm light generated by a combination of solid-state and fiber lasers with frequency mixing. We believe this laser will be beneficial for the application of interference lithography by seeding an injection-locking ArF eximer laser.

Single frequency, 5 ns, 200 µJ, 1553 nm fiber laser using silica based Er-doped fiber.

A 1553 nm Er-doped fiber master-oscillator-power-amplifier (MOPA) laser system providing pulses with a 6 kHz repetition rate, 5 ns duration, ~210 µJ energy, ~300 MHz linewidth, and with a near diffraction limited beam quality, was developed. A gain fiber as short as 0.7 m in length was utilized in order to relax the SBS effect. To the best of our knowledge, thus generated peak power of 40 kW is the highest one obtained from a single frequency Er-doped silica fiber laser. The pulse quality was verified by frequency conversion with a periodically poled lithium niobate nonlinear crystal (PPLN) for second harmonic generation. A pulse energy as high as ~100 µJ was achieved at 776.6 nm with a moderate incident energy of 133 µJ, indicating an energy conversion efficiency of 75%.

Hotspot Cancer Mutation Impairs KAT8-mediated Nucleosomal Histone Acetylation.

KAT8 is an evolutionarily conserved lysine acetyltransferase that catalyzes histone acetylation at H4K16 or H4K5 and H4K8 through distinct protein complexes. It plays a pivotal role in male X chromosome dosage compensation in Drosophila and is implicated in the regulation of diverse cellular processes in mammals. Mutations and dysregulation of KAT8 have been reported in human neurodevelopmental disorders and various cancers. However, the precise mechanisms by which these mutations disrupt KAT8's normal function, leading to disease pathogenesis, remain largely unknown. In this study, we focus on a hotspot missense cancer mutation, the R98W point mutation within the Tudor-knot domain. Our study reveals that the R98W mutation leads to a reduction in global H4K16ac levels in cells and downregulates the expression of target genes. Mechanistically, we demonstrate that R98 is essential for KAT8-mediated acetylation of nucleosomal histones by modulating substrate accessibility.

The Tudor-knot Domain of KAT5 Regulates Nucleosomal Substrate Acetylation.

The lysine acetyltransferase KAT5 is a pivotal enzyme responsible for catalyzing histone H4 acetylation in cells. In addition to its indispensable HAT domain, KAT5 also encompasses a conserved Tudor-knot domain at its N-terminus. However, the function of this domain remains elusive, with conflicting findings regarding its role as a histone reader. In our study, we have employed a CRISPR tiling array approach and unveiled the Tudor-knot motif as an essential domain for cell survival. The Tudor-knot domain does not bind to histone tails and is not required for KAT5's chromatin occupancy. However, its absence leads to a global reduction in histone acetylation, accompanied with genome-wide alterations in gene expression that consequently result in diminished cell viability. Mechanistically, we find that the Tudor-knot domain regulates KAT5's HAT activity on nucleosomes by fine-tuning substrate accessibility. In summary, our study uncovers the Tudor-knot motif as an essential domain for cell survival and reveals its critical role in modulating KAT5's catalytic efficiency on nucleosome and KAT5-dependent transcriptional programs critical for cell viability.

A potent and selective ENL degrader suppresses oncogenic gene expression and leukemia progression.

The histone acylation reader eleven-nineteen leukemia (ENL) plays a pivotal role in sustaining oncogenesis in acute leukemias, particularly in mixed-lineage leukemia-rearranged (MLL-r) leukemia. ENL relies on its reader domain to recognize histone lysine acylation promoting oncogenic gene expression and leukemia progression. Here, we report the development of MS41, a highly potent and selective von Hippel-Lindau-recruiting ENL degrader that effectively inhibits the growth of ENL-dependent leukemia cells. MS41-induced ENL degradation reduces the chromatin occupancy of ENL-associated transcription elongation machinery, resulting in the suppression of key oncogenic gene expression programs and the activation of differentiation genes. MS41 is well-tolerated in vivo and substantially suppresses leukemia progression in a xenograft mouse model of MLL-r leukemia. Notably, MS41 also induces the degradation of mutant ENL proteins identified in Wilms' tumors. Our findings emphasize the therapeutic potential of pharmacological ENL degradation for treating ENL-dependent cancers, making MS41 not only a valuable chemical probe but also potential anticancer therapeutic for further development.

TAZ2 truncation confers overactivation of p300 and cellular vulnerability to HDAC inhibition.

The histone acetyltransferase p300/CBP is composed of several conserved domains, among which, the TAZ2 domain is known as a protein-protein interaction domain that binds to E1A and various transcription factors. Here we show that TAZ2 has a HAT autoinhibitory function. Truncating p300/CBP at TAZ2 leads to hyperactive HAT and elevated histone H3K27 and H3K18 acetylation in cells. Mechanistically, TAZ2 cooperates with other HAT neighboring domains to maintain the HAT active site in a 'closed' state. Truncating TAZ2 or binding of transcription factors to TAZ2 induces a conformational change that 'opens' the active site for substrate acetylation. Importantly, genetic mutations that lead to p300/CBP TAZ2 truncations are found in human cancers, and cells with TAZ2 truncations are vulnerable to histone deacetylase inhibitors. Our study reveals a function of the TAZ2 domain in HAT autoinhibitory regulation and provides a potential therapeutic strategy for the treatment of cancers harboring p300/CBP TAZ2 truncations.

Molecular basis for nuclear accumulation and targeting of the inhibitor of apoptosis BIRC2.

The inhibitor of apoptosis protein BIRC2 regulates fundamental cell death and survival signaling pathways. Here we show that BIRC2 accumulates in the nucleus via binding of its second and third BIR domains, BIRC2BIR2 and BIRC2BIR3, to the histone H3 tail and report the structure of the BIRC2BIR3-H3 complex. RNA-seq analysis reveals that the genes involved in interferon and defense response signaling and cell-cycle regulation are most affected by depletion of BIRC2. Overexpression of BIRC2 delays DNA damage repair and recovery of the cell-cycle progression. We describe the structural mechanism for targeting of BIRC2BIR3 by a potent but biochemically uncharacterized small molecule inhibitor LCL161 and demonstrate that LCL161 disrupts the association of endogenous BIRC2 with H3 and stimulates cell death in cancer cells. We further show that LCL161 mediates degradation of BIRC2 in human immunodeficiency virus type 1-infected human CD4+ T cells. Our findings provide mechanistic insights into the nuclear accumulation of and blocking BIRC2.

TRIM28 secures skeletal stem cell fate during skeletogenesis by silencing neural gene expression and repressing GREM1/AKT/mTOR signaling axis.

Long bones are generated by mesoderm-derived skeletal progenitor/stem cells (SSCs) through endochondral ossification, a process of sequential chondrogenic and osteogenic differentiation tightly controlled by the synergy between intrinsic and microenvironment cues. Here, we report that loss of TRIM28, a transcriptional corepressor, in mesoderm-derived cells expands the SSC pool, weakens SSC osteochondrogenic potential, and endows SSCs with properties of ectoderm-derived neural crest cells (NCCs), leading to severe defects of skeletogenesis. TRIM28 preferentially enhances H3K9 trimethylation and DNA methylation on chromatin regions more accessible in NCCs; loss of this silencing upregulates neural gene expression and enhances neurogenic potential. Moreover, TRIM28 loss causes hyperexpression of GREM1, which is an extracellular signaling factor promoting SSC self-renewal and SSC neurogenic potential by activating AKT/mTORC1 signaling. Our results suggest that TRIM28-mediated chromatin silencing establishes a barrier for maintaining the SSC lineage trajectory and preventing a transition to ectodermal fate by regulating both intrinsic and microenvironment cues.

Immunosuppression Induced by Brain-Specific HDAC6 Knockdown Improves Aging Performance in Drosophila melanogaster.

HDAC6 is involved in several biological processes related to aging-associated diseases. However, it was unknown whether HDAC6 could directly regulate lifespan and healthspan. We found that HDAC6 knockdown induced transcriptome changes to attenuate the aging changes in the Drosophila head, particularly on the inflammation and innate immunity-related genes. Whole-body knockdown of HDAC6 extended lifespan in the fly, furthermore brain-specific knockdown of HDAC6 extended both lifespan and healthspan in the fly. Our results established HDAC6 as a lifespan regulator and provided a potential anti-aging target. Supplementary Information: The online version contains supplementary material available at 10.1007/s43657-022-00045-2.

Ultrafast Drift Current Terahertz Emission Amplification in the Monolayer WSe2/Si Heterostructure.

Two-dimensional transition metal dichalcogenides (TMDs) have great potential application for seamless on-chip integration due to their strong photon-electron-spin-valley coupling. However, the contact-free measurements of the valley-coupled photocurrent in TMDs is still challenging. Here, ultrafast terahertz emission spectroscopy is employed to investigate the photocurrent dynamics in monolayer WSe2, and an interface-induced drift current amplification is found in the WSe2/Si heterostructure. The amplification of terahertz emission comes from the photocurrent enlarged by band bending in the WSe2 and Si junction, and the amplification ratio increase further near the valley resonant transition of WSe2. In addition, the valley-momentum locked photocurrent in the WSe2/Si heterostructure reserves the same chirality with monolayer WSe2 at room temperature. These findings could provide a new method for manipulating valley-momentum locked photocurrent by photon helicity and open new avenues for TMD-based valley-polarized terahertz emission devices.