Rab32 promotes glioblastoma migration and invasion via regulation of ERK/Drp1-mediated mitochondrial fission.

The highly widespread and infiltrative nature of glioblastoma multiforme (GBM) makes complete surgical resection hard, causing high recurrence rate and poor patients' prognosis. However, the mechanism underlying GBM migration and invasion is still unclear. In this study, we investigated the role of a Ras-related protein Rab32 on GBM and uncovered its underlying molecular and subcellular mechanisms that contributed to GBM aggressiveness. The correlation of Rab32 expression with patient prognosis and tumor grade was investigated by public dataset analysis and clinical specimen validation. The effect of Rab32 on migration and invasion of GBM had been evaluated using wound healing assay, cell invasion assay, as well as protein analysis upon Rab32 manipulations. Mitochondrial dynamics of cells upon Rab32 alterations were detected by immunofluorescence staining and western blotting. Both the subcutaneous and intracranial xenograft tumor model were utilized to evaluate the effect of Rab32 on GBM in vivo. The expression level of Rab32 is significantly elevated in the GBM, especially in the most malignant mesenchymal subtype, and is positively correlated with tumor pathological grade and poor prognosis. Knockdown of Rab32 attenuated the capability of GBM's migration and invasion. It also suppressed the expression levels of invasion-related proteins (MMP2 and MMP9) as well as mesenchymal transition markers (N-cadherin, vimentin). Interestingly, Rab32 transported Drp1 to mitochondrial from the cytoplasm and modulated mitochondrial fission in an ERK1/2 signaling-dependent manner. Furthermore, silencing of Rab32 in vivo suppressed tumor malignancy via ERK/Drp1 axis. Rab32 regulates ERK1/2/Drp1-dependent mitochondrial fission and causes mesenchymal transition, promoting migration and invasion of GBM. It serves as a novel therapeutic target for GBM, especially for the most malignant mesenchymal subtype. Schematic of Rab32 promotes GBM aggressiveness via regulation of ERK/Drp1-mediated mitochondrial fission. Rab32 transports Drp1 from the cytoplasm to the mitochondria and recruits ERK1/2 to activate the ser616 site of Drp1, which in turn mediates mitochondrial fission and promotes mesenchymal transition, migration and invasion of GBM.

3D printing of dynamic covalent polymer network with on-demand geometric and mechanical reprogrammability.

Delicate geometries and suitable mechanical properties are essential for device applications of polymer materials. 3D printing offers unprecedented versatility, but the geometries and mechanical properties are typically fixed after printing. Here, we report a 3D photo-printable dynamic covalent network that can undergo two independently controllable bond exchange reactions, allowing reprogramming the geometry and mechanical properties after printing. Specifically, the network is designed to contain hindered urea bonds and pendant hydroxyl groups. The homolytic exchange between hindered urea bonds allows reconfiguring the printed shape without affecting the network topology and mechanical properties. Under different conditions, the hindered urea bonds are transformed into urethane bonds via exchange reactions with hydroxyl groups, which permits tailoring of the mechanical properties. The freedom to reprogram the shape and properties in an on-demand fashion offers the opportunity to produce multiple 3D printed products from one single printing step.

Miniature Noninvasive Sensor Based on Impedance-Change Detection in Branches for Measuring Branch Ice Content in Overwintering Woody Plants.

Advancements in detection instruments have enabled the real-time acquisition of water information during plant growth; however, the real-time monitoring of freeze-thaw information during plant overwintering remains a challenge. Based on the relationship between the change in the water-ice ratio and branch impedance during freezing, a miniature noninvasive branch volume ice content (BVIC) sensor was developed for monitoring real-time changes in volumetric ice content and the ice freeze-thaw rate of woody plant branches during the overwintering period. The results of the performance analysis of the impedance measurement circuit show that the circuit has a lateral sensitivity range, measurement range, resolution, measurement accuracy, and power consumption of 0-35 mm, 0-100%, 0.05%, ±1.76%, and 0.25 W, respectively. The dynamic response time was 0.296 s. The maximum allowable error by the output voltage fluctuation, owing to the ambient temperature and humidity, was only ±0.635%, which meets the actual use requirements. The calibration curve fit coefficients were >0.98, indicating a significant correlation. The ice content of plant branches under cold stress was measured for indoor and field environments, and the sensors could effectively monitor changes in the branch ice content in plants exposed to cold stress. Additionally, they can differentiate between plants with different cold resistances, indicating the reliability of the BVIC sensor.

A Novel Bird-Shape Broadband Piezoelectric Energy Harvester for Low Frequency Vibrations.

This work presents a novel bird-shaped broadband piezoelectric energy harvester based on a two-DOF crossed beam for low-frequency environmental vibrations. The harvester features a cantilever mounted on a double-hinged beam, whose rotating motions effectively diminish its natural frequencies. Numerical simulation based on the finite element method is conducted to analyze the modal shapes and the harmonic response of the proposed harvester. Prototypes are fabricated and experiments are carried out by a testing system, whose results indicate a good agreement with the simulation. The multi-frequency energy harvesting is achieved at the first-, second-, and fifth-order resonances. In particular, the proposed harvester demonstrates the remarkable output characteristics of 9.53 mW and 1.83 mW at frequencies as low as 19.23 HZ and 45.38 Hz, which are superior to the majority of existing energy harvesters. Besides, the influences of key parameters on the harvesting performance are experimentally investigated to optimize the environmental adaptability of the harvester. This work provides a new perspective for efficiently harvesting the low-frequency vibration energy, which can be utilized for supplying power to electronic devices.

Multi-omics analysis of N6-methyladenosine reader IGF2BP3 as a promising biomarker in pan-cancer.

Background: Insulin-like growth factor 2 mRNA-binding protein 3 (IGF2BP3) has been reported to exhibit an oncogenic effect as an RNA-binding protein (RBP) by promoting tumor cell proliferation, migration and invasion in several tumor types. However, a pan-cancer analysis of IGF2BP3 is not currently available, and the exact roles of IGF2BP3 in prognosis and immunology in cancer patients remain enigmatic. The main aim of this study was to provide visualization of the systemic prognostic landscape of IGF2BP3 in pan-cancer and to uncover the potential relationship between IGF2BP3 expression in the tumor microenvironment and immune infiltration profile. Methods: Raw data on IGF2BP3 expression were obtained from GTEx, CCLE, TCGA, and HPA data portals. We have investigated the expression patterns, diagnostic and prognostic significance, mutation landscapes, functional analysis, and functional states of IGF2BP3 utilizing multiple databases, including HPA, TISIDB, cBioPortal, GeneMANIA, GESA, and CancerSEA. Moreover, the relationship of IGF2BP3 expression with immune infiltrates, TMB, MSI and immune-related genes was evaluated in pan-cancer. IGF2BP3 with drug sensitivity analysis was performed from the CellMiner database. Furthermore, the expression of IGF2BP3 in different grades of glioma was detected by immunohistochemical staining and western blot. Results: We found that IGF2BP3 was ubiquitously highly expressed in pan-cancer and significantly correlated with diagnosis, prognosis, TMB, MSI, and drug sensitivity in various types of cancer. Besides, IGF2BP3 was involved in many cancer pathways and varied in different immune and molecular subtypes of cancers. Additionally, IGF2BP3 is critically associated with genetic markers of immunomodulators in various cancers. Finally, we validated that IGF2BP3 protein expression was significantly higher in glioma than in normal tissue, especially in GBM. Conclusions: IGF2BP3 may be a potential molecular biomarker for diagnosis and prognosis in pan-cancer, especially for glioma. It could become a novel therapeutic target for various cancers.

Aminopeptidase N-Responsive Conjugates with Tunable Charge-Reversal Properties for Highly Efficient Tumor Accumulation and Penetration.

Tumor enzyme-responsive charge-reversal carriers can induce efficient transcytosis and lead to efficient tumor infiltration and potent anticancer efficacy. However, the correlations of molecular structure with charge-reversal property, tumor penetration, and drug delivery efficiency are unknown. Herein, aminopeptidase N (APN)-responsive conjugates were synthesized to investigate these correlations. We found that the monomeric unit structure and the polymer chain structure determined the enzymatic hydrolysis and charge-reversal rates, and accordingly, the transcytosis and tumor accumulation and penetration of the APN-responsive conjugates. The conjugate with moderate APN responsiveness balanced the in vitro transcytosis and in vivo overall drug delivery process and achieved the best tumor delivery efficiency, giving potent antitumor efficacy. This work provides new insight into the design of tumor enzyme-responsive charge-reversal nanomedicines for efficient cancer drug delivery.

Regenerative Living 4D Printing via Reversible Growth of Polymer Networks.

Living creatures possess complex geometries, exceptional adaptability, and continuous growing and regenerating characteristics, which are difficult for synthetic materials to imitate simultaneously. A living polymer network with these features is reported. The polymer can be digitally printed into arbitrary 3D shapes and subsequently undergoes growth via living polymerization of a monomer as the nutrient. This leads to macroscopic dimensional growth and transforms the printed amorphous network into a crystallizable network, resulting in geometric adaptability via a shape-memory mechanism. By controlling the localized growth, an initial homogeneous structure can be converted into a geometrically different heterogeneous structure composed of materials with different properties (crystallization and mechanical properties). After growth, the original network can be chemically regenerated for regrowth. With this regenerative living 4D printing, one 3D-printed seed template can be turned into different derivatives with distinct geometries and mechanical properties when repeated regeneration is conducted in different localized regions and the degree of regrowth is varied.

Orthogonal Photochemistry toward Direct Encryption of a 3D-Printed Hydrogel.

Encryption technologies are essential for information security and product anti-counterfeiting, but they are typically restricted to planar surfaces. Encryption on complex 3D objects offers great potential to further improve security. However, it is rarely achieved owing to the lack of encoding strategies for nonplanar surfaces. Here, an approach is reported to directly encrypt on a 3D-printed object employing orthogonal photochemistry. In this system, visible light photochemistry is used for 3D printing of a hydrogel, and ultraviolet light is subsequently employed to activate its geometrically complex surface through the dissociation of ortho-nitrobenzyl ester units in a spatioselective manner for information coding. This approach offers a new way for more reliable encryption, and the underlying orthogonal photochemistry can be extended toward functional modification of 3D-printed products beyond information protection.

Proteogenomics of diffuse gliomas reveal molecular subtypes associated with specific therapeutic targets and immune-evasion mechanisms.

Diffuse gliomas are devastating brain tumors. Here, we perform a proteogenomic profiling of 213 retrospectively collected glioma tumors. Proteogenomic analysis reveals the downstream biological events leading by EGFR-, IDH1-, TP53-mutations. The comparative analysis illustrates the distinctive features of GBMs and LGGs, indicating CDK2 inhibitor might serve as a promising drug target for GBMs. Further proteogenomic integrative analysis combined with functional experiments highlight the cis-effect of EGFR alterations might lead to glioma tumor cell proliferation through ERK5 medicates nucleotide synthesis process. Proteome-based stratification of gliomas defines 3 proteomic subgroups (S-Ne, S-Pf, S-Im), which could serve as a complement to WHO subtypes, and would provide the essential framework for the utilization of specific targeted therapies for particular glioma subtypes. Immune clustering identifies three immune subtypes with distinctive immune cell types. Further analysis reveals higher EGFR alteration frequencies accounts for elevation of immune check point protein: PD-L1 and CD70 in T-cell infiltrated tumors.

Using Downgaze Palsy Progression Rate to Model Survival in Progressive Supranuclear Palsy-Richardson Syndrome.

BACKGROUND: Rapid development of downgaze palsy, the most specific symptom of progressive supranuclear palsy (PSP), has been associated with shorter survival in small studies. OBJECTIVE: We hypothesized that the progression rate of downgaze palsy and other disease features could predict survival if assessed soon after the onset of downgaze palsy in a large data set. METHODS: We used a longitudinal database of 414 patients with probable PSP-Richardson syndrome from 1994 to 2020. The data set comprised demographics and, for each visit, 28 PSP Rating Scale (PSPRS) items and PSP stage scores. We calculated the rate of progression of each PSPRS item as its item score when the downgaze item first reached 1 or more (on a 0-4 scale) divided by disease duration at that point. Multivariate Cox regression was applied to identify variables independently associated with survival. We also explored the progression pattern of total PSPRS and downgaze palsy scores with disease course. RESULTS: Independently associated with shorter survival were older onset age and faster progression of downgaze palsy, dysphagia for liquids, difficulty in returning to seat, and PSP stage. Patients with survival duration within 1 year of the median survival (6.58 years) showed approximately linear progression of the PSPRS score and downgaze palsy score during years 2 through 6 of the disease course. CONCLUSIONS: Older onset age and faster progression of downgaze palsy and several axial features are associated with shorter survival. The disease typically progresses in approximately linear fashion during years 2 through 6. These results may aid study design and patient counseling. © 2022 The Authors. Movement Disorders published by Wiley Periodicals LLC on behalf of International Parkinson and Movement Disorder Society.

Osthole increases the radiosensitivity of hepatoma cells by inhibiting GSK-3ß/AMPK/mTOR pathway-controlled glycolysis.

Osthole is a natural coumarin substance that has an inhibitory effect on hepatic cancer, but its radiosensitization effect on hepatoma cells has not been reported. This study aimed to investigate the effect of osthole. Human HCC-LM3 and SK-Hep-1 hepatoma cells were used and treated with or without osthole, irradiation, or their combination; the cell survival, migration, colony formation, DNA damage repair, intracellular lactic acid content, and glycolysis-related glycogen synthase kinase-3ß (GSK-3ß), p-GSK-3ß, AMP-activated protein kinase (AMPK), p-AMPK, mammalian target of rapamycin (mTOR), p-mTOR, glucose transporter-1 (GLUT-1), GLUT-3, and pyruvate kinase isozyme type M2 (PKM2) protein expressions were determined. Compared with the irradiation group, the osthole plus irradiation group could further decrease the survival rate, migration, colony formation, and DNA damage repair of both hepatoma cells, indicating a synergistic effect of the combination treatment. Moreover, the combination of osthole and irradiation could decrease the content of intracellular lactic acid, ratios of intracellular p-GSK-3ß/GSK-3ß and p-mTOR/mTOR proteins, and expressions of intracellular GLUT-1/3 and PKM2 proteins, and increase the ratio of intracellular p-AMPK/AMPK proteins. Osthole can increase the radiosensitivity of hepatoma cells, and its radiosensitization mechanisms may be related to glycolytic inhibition by attenuating the GSK-3ß/AMPK/mTOR pathway.

An R3-MYB repressor, BnCPC forms a feedback regulation with MBW complex to modulate anthocyanin biosynthesis in Brassica napus.

BACKGROUND: Anthocyanins are metabolites of phenylpropanoid pathway, and involves in diverse processes of plant development and adaptation, which are regulated by the MYB-bHLH-WD40 (MBW) protein complexes. Many R2R3-MYB activators have been well characterized, but the MYB repressors in anthocyanin biosynthesis were recognized recently, which are also important in modulating phenylpropanoid metabolism in plants. The regulatory mechanism of anthocyanin biosynthesis in oil crop Brassica napus remains to be revealed. RESULTS: In this study, we identified an anthocyanin repressor BnCPC in B. napus. BnCPC encoded a typical R3-MYB protein containing a conserved [D/E]Lx2[R/K]x3Lx6Lx3R motif for interaction with bHLH proteins. Overexpression of BnCPC in B. napus inhibited anthocyanin accumulation, especially under anthocyanin inducible conditions. Protein-protein interaction and dual-luciferase assays confirmed that BnCPC could compete with BnPAP1 to interact with bHLHs (BnTT8 and BnEGL3), and repress the expression of anthocyanin biosynthetic genes (e.g., BnDFR) that activated by MBW complexes. Moreover, we found BnCPC inhibited the MBW complex-induced BnCPC activity. CONCLUSIONS: Overall, this research demonstrated that BnCPC repressed anthocyanin biosynthesis by affecting the formation of MBW complex, and formed a feedback loop to regulate anthocyanin accumulation in B. napus.

Effects of mixed-cooling strategies on executive functions in simulated tennis in hot and humid conditions.

This study aimed to investigate the effects of mixed-cooling strategies, which combines external (cooling vest + neck cooled collar) and internal cooling (cold sports drink ingestion) on measures of executive function during simulated tennis in hot/humid conditions. In a counterbalanced design (randomised order), eight males undertook two trials [one with the mixed-cooling strategy, (MCOOL condition) and another without (CON condition)] in a climate chamber (36.5°C, 50% relative humidity). All subjects completed an intermittent treadmill protocol simulating a three-set tennis match with a 90-second break during odd-numbered games and 120-second breaks between sets, in accordance with the activity profile and International Tennis Federation rules. The mixed-cooling strategies were adopted before test and break time during the simulated tennis match. Stroop task, 2-back task, More-odd shifting task, gastrointestinal temperature (Tgi), skin temperature, blood lactic acid (BLA), heart rate, urine specific gravity (USG), sweat rate (SR), thermal sensation (TS) and perceived exertion (RPE) were measured. Results showed that the mean exercise time was longer in the MCOOL condition than in the CON condition. The SR was greater in CON trial compared with that in MCOOL trial. Results of two-way analysis of variance with repeated measures revealed that time×condition interactions were significant in BLA, Stroop response time, and switch cost of the more-odd shifting task. There were main effects of condition for Tgi, HR, TS, RPE, BLA, Stroop response time, and switch cost of the more-odd shifting task. In a hot/wet environment, pre- and intermittent mixed-cooling strategies can significantly improve exercise time and measures of executive function of tennis players in a simulated tennis match.

Endoscopic transcortical expanded transforaminal transvenous transchoroidal approach to third ventricle lesion resection using an endoport.

OBJECTIVE: To investigate the clinical experience and application value of endoscopic resection of lesions in and around the third ventricle using a transcortical expanded transforaminal transvenous transchoroidal approach with an endoport. METHODS: Clinical data and follow-up results of seven patients who underwent the removal of lesions in the third ventricle and its adjacent area with an endoport-guided endoscopic system from January 2018 to December 2020 in the Department of Neurosurgery, Zhongshan Hospital Affiliated to Fudan University, were analyzed retrospectively. Two other patients from the Affiliated Pediatric Hospital of Fudan University and the Affiliated Hospital of Guizhou Medical University, respectively, were included in the analysis. RESULTS: A total of nine cases of third ventricle tumors were included in the study, including six women and three men, with an average age of 37.8 years (4-84 years old) and a follow-up time of 6-44 months. These nine tumor cases included two pilocytic astrocytomas, one diffuse midline glioma (H3 K27-altered), two craniopharyngiomas, two choroid plexus (CP) papillomas, one germinoma, and one pineal parenchymal tumor of intermediate differentiation. Total resection was completed in eight cases, with one near-total resection. There were no complications related to the surgical approach, such as epilepsy, aphasia, or hemiplegia. CONCLUSIONS: The endoscope transcortical expanded transforaminal transvenous transchoroidal approach using an endoport can safely and effectively remove third ventricle lesions. This approach can reach a wide area, from the anterior to the posterior third ventricle.

[Professor ZHANG Jian-bin's clinical experience in "moving cupping therapy on the back"].

This paper summarizes Professor ZHANG Jian-bin's experience in "moving cupping therapy on the back" in clinical practice. Professor ZHANG Jian-bin is good at applying the theory of governor vessel to the clinical diagnosis and treatment. He believes that the moving cupping therapy is the supplementation of "spinal therapy" in clinical diagnosis and treatment. He emphasizes that during the moving cupping therapy exerted, the physicians should observe carefully to in-time obtain the perceived feedback of treatment in patients. Afterward, the cupping marks left should be inspected to discover the location of disorder in the body, thus, a targeted treatment can be provided. Besides, the prognosis of the disease can be judged by the feedback after many treatments with the moving cupping therapy. The moving cupping therapy displays its important value in the diagnosis and treatment of disease, as well as the evaluation of the prognosis.

Endoscopic resection of thalamic lesions via supracerebellar infratentorial approach: a case series and technical note.

The deep location of the thalamus and the complex neural circuits in the surrounding area make surgery extremely challenging. Feasibility and advisability of using a supracerebellar infratentorial approach (SCITA) for endoscopic resection of thalamic lesions remains to be further evaluated. Fifteen patients who underwent endoscopic resection of thalamic via SCITA from 2014 to 2021 were retrospectively collected. We analyzed preoperative tumor-related variables and surgical procedures in detail, as well as postoperative outcomes. Lesions mainly located in the posterior and/or medial part of the thalamus, and some of them expanded downward, or backward. The mean size of them was 30 × 24 mm. Five of the nine patients with preoperative hydrocephalus underwent cerebrospinal fluid shunts to relieve increased intracranial pressure. Among the 15 patients, 4 were glioblastoma, 3 were pilocytic astrocytoma, and the rest included 1 case of anaplastic astrocytoma, melanoma, polymorphous low-grade neuroepithelial tumor of the young, rosette-forming glioneuronal tumor, inflammatory lesion, diffuse midline glioma, and cavernous hemangioma. The majority of patients (10/15) achieved gross total resection, which has become more pronounced since paramedian SCITA was used in place of the midline approach in 2020 (6/8). Three patients had unresolved or new onset of clinical symptoms after surgery, resulting in a decreased KPS score at discharge. Neuro-endoscopic techniques can ameliorate many of the shortcomings of the SCITA. With the accumulation of experience and technological progress, more deficiencies of this approach may be improved, enabling safe and effective resection of posterior and/or medial part thalamic lesions.

The Aurantii Fructus Immaturus flavonoid extract alleviates inflammation and modulate gut microbiota in DSS-induced colitis mice.

Inflammatory bowel disease (IBD) is a chronic, relapsing immune-mediated disease that always leads to a progressive loss of intestinal function. Therefore, it is important to find potential therapeutic drugs. This study was conducted to elucidate the effect of Aurantii Fructus immaturus flavonoid extract (AFI, 8% neohesperidin, 10% naringin) on DSS-induced intestinal inflammation and the gut microbiome. To explore the mechanism of action by which AFI protects against intestinal inflammation, a total of 50 mice were randomly divided into 5 groups [CG (control group), MG (model group), AFI low dose, AFI middle dose, and AFI high dose] and received 2.5% DSS for 7 days. Then, mice in the AFI groups were orally administered different doses of AFI for 16 days. The results showed that, compared with the MG group, the food intake and body weight were increased in the AFI groups, but the water intake was lower. Additionally, AFI significantly alleviated DSS-induced colitis symptoms, including disease activity index (DAI), and colon pathological damage. The levels of IL-6, IL-1ß and TNF-α in serum and colon tissue were significantly decreased. The diversity and abundance of the intestinal microbiota in the AFI group were decreased. The relative abundance of Bacteroidota was increased, and the relative abundance of Firmicutes was decreased. AFI plays an important role in alleviating DSS-induced intestinal inflammation and regulating Oscillospira, Prevotellaceae and Lachnospiraceae in the intestine at low, medium and high doses, respectively. This report is a pioneer in the assessment of AFI. This study not only demonstrated the anti-inflammatory activity of AFI but also identified the microbiota regulated by different concentrations of AFI.

Dielectric Polymer with Designable Large Motion under Low Electric Field.

Dielectric elastomers (DEs) can demonstrate fast and large in-plane expansion/contraction due to electric field (e-field)-induced Maxwell stress. For robotic applications, it is often necessary that the in-plane actuation is converted into out-of-plane motions with mechanical frames. Despite their performance appeal, their high driving e-field (20-100 V µm-1 ) demands bulky power accessories and severely compromises their durability. Here, a dielectric polymer that can be programmed into diverse motions actuated under a low e-field (2-10 V µm-1 ) is reported. The material is a crystalline dynamic covalent network that can be reconfigured into arbitrary 3D geometries. This gives rise to a geometric effect that markedly amplifies the actuation, leading to designable large motions when the dielectric polymer is heated above its melting temperature to become a DE. Additionally, the crystallization transition enables dynamic multimodal motions and active deployability. These attributes result in unique design versatility for soft robots.

Targeting Homologous Recombination Deficiency in Ovarian Cancer with PARP Inhibitors: Synthetic Lethal Strategies That Impact Overall Survival.

The advent of molecular targeted therapies has made a significant impact on survival of women with ovarian cancer who have defects in homologous recombination repair (HRR). High-grade serous ovarian cancer (HGSOC) is the most common histological subtype of ovarian cancer, with over 50% displaying defective HRR. Poly ADP ribose polymerases (PARPs) are a family of enzymes that catalyse the transfer of ADP-ribose to target proteins, functioning in fundamental cellular processes including transcription, chromatin remodelling and DNA repair. In cells with deficient HRR, PARP inhibitors (PARPis) cause synthetic lethality leading to cell death. Despite the major advances that PARPis have heralded for women with ovarian cancer, questions and challenges remain, including: can the benefits of PARPis be brought to a wider range of women with ovarian cancer; can other drugs in clinical use function in a similar way or with greater efficacy than currently clinically approved PARPis; what can we learn from long-term responders to PARPis; can PARPis sensitise ovarian cancer cells to immunotherapy; and can synthetic lethal strategies be employed more broadly to develop new therapies for women with ovarian cancer. We examine these, and other, questions with focus on improving outcomes for women with ovarian cancer.