Neurosymptomatic HIV-1 CSF escape is associated with replication in CNS T cells and inflammation.

During antiretroviral therapy (ART), most people living with HIV-1 have undetectable HIV-1 RNA in their plasma. However, they occasionally present with new or progressive neurologic deficits and detectable HIV-1 RNA in the cerebrospinal fluid (CSF), a condition defined as neurosymptomatic HIV-1 CSF escape (NSE). We explored the source of neuropathogenesis and HIV-1 RNA in the CSF during NSE by characterizing HIV-1 populations and inflammatory biomarkers in CSF from 25 individuals with NSE. HIV-1 populations in the CSF were uniformly drug resistant and adapted to replication in CD4+ T cells, but differed greatly in genetic diversity, with some having low levels of diversity similar to those observed during untreated primary infection and others having high levels like those during untreated chronic infection. Higher diversity in the CSF during NSE was associated with greater CNS inflammation. Finally, optimization of ART regimen was associated with viral suppression and improvement of neurologic symptoms. These results are consistent with CNS inflammation and neurologic injury during NSE being driven by replication of partially drug-resistant virus in CNS CD4+ T cells. This is unlike nonsuppressible viremia in the plasma during ART, which typically lacks clinical consequences and is generated by virus expression without replication.

Changes in cerebrospinal fluid proteins across the spectrum of untreated and treated chronic HIV-1 infection.

Using the Olink Explore 1536 platform, we measured 1,463 unique proteins in 303 cerebrospinal fluid (CSF) specimens from four clinical centers contributed by uninfected controls and 12 groups of people living with HIV-1 infection representing the spectrum of progressive untreated and treated chronic infection. We present three initial analyses of these measurements: an overview of the CSF protein features of the sample; correlations of the CSF proteins with CSF HIV-1 RNA and neurofilament light chain protein (NfL) concentrations; and comparison of CSF proteins in HIV-associated dementia (HAD) and neurosymptomatic CSF escape (NSE). These reveal a complex but coherent picture of CSF protein changes with highest concentrations of many proteins during CNS injury in the HAD and NSE groups and variable protein changes across the course of systemic HIV-1 progression that included two common patterns, designated as lymphoid and myeloid patterns, related to principal involvement of their underlying inflammatory cell lineages. Antiretroviral therapy reduced CSF protein perturbations, though not always to control levels. The dataset of these CSF protein measurements, along with background clinical information, is posted online. Extended studies of this unique dataset will supplement this report to provide more detailed characterization of the dynamic impact of HIV-1 infection on the CSF proteome across the spectrum of HIV-1 infection, advancing the mechanistic understanding of HIV-1-related CNS pathobiology.

Bendable non-silicon RISC-V microprocessor.

Semiconductors have already had a very profound effect on society, accelerating scientific research and driving greater connectivity. Future semiconductor hardware will open up new possibilities in quantum computing, artificial intelligence and edge computing, for applications such as cybersecurity and personalized healthcare. By nature of its ethos, open hardware provides opportunities for even greater collaboration and innovations across education, academic research and industry. Here we present Flex-RV, a 32-bit microprocessor based on an open RISC-V (ref. 1) instruction set fabricated with indium gallium zinc oxide thin-film transistors2 on a flexible polyimide substrate, enabling an ultralow-cost bendable microprocessor. Flex-RV also integrates a programmable machine learning (ML) hardware accelerator inside the microprocessor and demonstrates new instructions to extend the RISC-V instruction set to run ML workloads. It is implemented, fabricated and demonstrated to operate at 60 kHz consuming less than 6 mW power. Its functionality when assembled onto a flexible printed circuit board is validated while executing programs under flat and tight bending conditions, achieving no worse than 4.3% performance variation on average. Flex-RV pioneers an era of sub-dollar open standard non-silicon 32-bit microprocessors and will democratize access to computing and unlock emerging applications in wearables, healthcare devices and smart packaging.

The Dark Art of Light Curing in Dentistry.

OBJECTIVE: This study was designed to show that the commonly reported irradiance values that are quoted in most publications are inadequate to describe the light output from light curing units (LCUs). METHODS: The total spectral radiant power (mW) output from 12 contemporary LCUs was measured with a fiberoptic spectroradiometer and a calibrated integrating sphere. Five recordings were taken for each LCU and exposure mode. In addition, the irradiances (mW/cm²) delivered at 0-mm, 5-mm and 10-mm distances were recorded through a 6-mm diameter aperture and radiant exposures (J/cm²) from the LCUs were calculated. Light beam profiles from the LCUs were recorded using a beam profiler, and the images were overlaid on a molar tooth to simulate a clinical setting. Data were analyzed using ANOVA followed by Tukey post-hoc test (α=0.05). RESULTS: The mean power outputs from the LCUs ranged from 380 to 2472 mW (p<0.0001). The highest irradiance was recorded from the Cicada CV 215-G7 (3091 mW/cm² in its highest mode) and the lowest from the Radii Cal CX (731 mW/cm²). The emission spectra differed, even among the multi-peak and single-peak LCUs. Radiant exposures from the entire light tip ranged from 18.3 J/cm², Radii Cal CX, in its standard 25 s exposure mode to 3.9 J/cm² from the Monet Laser in a 3 s exposure setting. Half (50%) of the measured irradiance values from the LCUs differed from the manufacturers' value by more than 10%. There were significant differences in the impact of distance from the tip. The beam profiles visually highlighted the varying effects of distance from the LCU tip among different units. CONCLUSION: There were significant differences in the emission spectra, power outputs, tip diameters, irradiances, radiant exposures, and the effect distance from the light tips. These differences underline the importance of manufacturers and researchers correctly measuring and reporting the output from the LCU to ensure that research is reproducible and that patients receive acceptable dental restorations.

Focused Ultrasound Impels the Delivery and Penetration of Model Therapeutics into Cerebral Cavernous Malformations.

BACKGROUND: Cerebral cavernous malformations (CCMs) are vascular neoplasms in the brain that can cause debilitating symptoms. Current treatments pose significant risks to some patients, motivating the development of new nonsurgical options. We recently discovered that focused ultrasound-mediated blood-brain barrier opening (FUS) arrests CCM formation and growth. Here, we build on this discovery and assess the ability of FUS to deliver model therapeutics into CCMs. METHODS: Quantitative T1 mapping MRI sequences were used with 1 kDa (MultiHance; MH) and 17 kDa (GadoSpin D; GDS) contrast agents to assess the FUS-mediated delivery and penetration of model small molecule drugs and biologics, respectively, into CCMs of Krit1 mutant mice. RESULTS: FUS elevated the rate of MH delivery to both the lesion core (4.6-fold) and perilesional space (6.7-fold). Total MH delivery more than doubled in the lesion core and tripled in the perilesional space when FUS was applied immediately prior to MH injection. For the model biologic drug (i.e. GDS), FUS was of greater relative benefit, resulting in 21.7-fold and 3.8-fold delivery increases to the intralesional and perilesional spaces, respectively. CONCLUSIONS: FUS is capable of impelling the delivery and penetration of therapeutics into the complex and disorganized CCM microenvironment. Benefits to small molecule drug delivery are more evident in the perilesional space, while benefits to biologic delivery are more evident in CCM cores. These findings, when combined with ability of FUS alone to control CCMs, highlight the potential of FUS to serve as a powerful non-invasive therapeutic platform for CCM.

Tissue- and Temporal-Dependent Dynamics of Myeloablation in Response to Gemcitabine Chemotherapy.

For triple-negative breast cancer (TNBC), the most aggressive subset of breast cancer, immune cell infiltrates have prognostic implications. The presence of myeloid-derived suppressor cells supports tumor progression, while tumor-infiltrating lymphocytes (TILs) correlate with improved survival and responsiveness to immunotherapy. Manipulating the abundance of these populations may enhance tumor immunity. Gemcitabine (GEM), a clinically employed chemotherapeutic, is reported to be systemically myeloablative, and thus it is a potentially useful adjunct therapy for promoting anti-tumor immunity. However, knowledge about the immunological effects of GEM intratumorally is limited. Thus, we directly compared the impact of systemic GEM on immune cell presence and functionality in the tumor microenvironment (TME) to its effects in the periphery. We found that GEM is not myeloablative in the TME; rather, we observed sustained, significant reductions in TILs and dendritic cells-crucial components in initiating an adaptive immune response. We also performed bulk-RNA sequencing to identify immunological alterations transcriptionally induced by GEM. While we found evidence of upregulation in the interferon-gamma (IFN-γ) response pathway, we determined that GEM-mediated growth control is not dependent on IFN-γ. Overall, our findings yield new insights into the tissue- and temporal-dependent immune ablative effects of GEM, contrasting the paradigm that this therapy is specifically myeloablative.

Brain Nucleic Acid Delivery and Genome Editing via Focused Ultrasound-Mediated Blood-Brain Barrier Opening and Long-Circulating Nanoparticles.

We introduce a two-pronged strategy comprising focused ultrasound (FUS)-mediated blood-brain barrier (BBB) opening and long-circulating biodegradable nanoparticles (NPs) for systemic delivery of nucleic acids to the brain. Biodegradable poly(ß-amino ester) polymer-based NPs were engineered to stably package various types of nucleic acid payloads and enable prolonged systemic circulation while retaining excellent serum stability. FUS was applied to a predetermined coordinate within the brain to transiently open the BBB, thereby allowing the systemically administered long-circulating NPs to traverse the BBB and accumulate in the FUS-treated brain region, where plasmid DNA or mRNA payloads produced reporter proteins in astrocytes and neurons. In contrast, poorly circulating and/or serum-unstable NPs, including the lipid NP analogous to a platform used in clinic, were unable to provide efficient nucleic acid delivery to the brain regardless of the BBB-opening FUS. The marriage of FUS-mediated BBB opening and the long-circulating NPs engineered to copackage mRNA encoding CRISPR-associated protein 9 and single-guide RNA resulted in genome editing in astrocytes and neurons precisely in the FUS-treated brain region. The combined delivery strategy provides a versatile means to achieve efficient and site-specific therapeutic nucleic acid delivery to and genome editing in the brain via a systemic route.

Evaluation of the information provided in the instruction manuals of dental light-curing units.

OBJECTIVE: This study evaluated the completeness and accuracy of information in LCU instruction manuals from 40 manufacturers. MATERIALS AND METHODS: Instruction manuals from 40 LCUs (20 from leading manufacturers and 20 budget units) were reviewed. Twenty-eight parameters across five categories were assessed using a binary scale (0=incorrect/missing, 1=correct). The categories and their respective evaluation scores were: LCU characteristics (43%), instructions for use (7%), safety precautions (14%), maintenance recommendations (29%), and regulatory certification (7%). These scores were combined to produce a final score. RESULTS: Scores from leading manufacturers ranged between 46-86%, while the budget category ranged from 18-68%. All manuals provided information about the wavelength/spectrum of the LCU. Only Valo X and Valo Cordless reported power values and used the term "irradiance" instead of "intensity." Details such as LED type and active tip emission area were often missing. Instructions on how to use the LCU to photo-cure resins were frequently limited. Although most manuals addressed safety precautions, several lacked details on heat issues and general health precautions. All manuals included maintenance instructions, though information on replacement parts was often missing. Among the LCUs, 85% stated they were CE certified, 32% held both FDA and CE certification, and 63% claimed compliance with ISO and/or IEC standards. CONCLUSIONS: There were notable differences in the completeness and accuracy of the instruction manuals. Manuals from major manufacturers generally provided more comprehensive information than their budget counterparts. CLINICAL SIGNIFICANCE: Instruction manuals should contain accurate information to help clinicians deliver the highest standard of care. The lack of important information about the LCUs in the manuals is concerning.

Changes in Cerebrospinal Fluid Proteins across the Spectrum of Untreated and Treated Chronic HIV-1 Infection.

Using the Olink Explore 1536 platform, we measured 1,463 unique proteins in 303 cerebrospinal fluid (CSF) specimens from four clinical centers that included uninfected controls and 12 groups of people living with HIV-1 infection representing the spectrum of progressive untreated and treated chronic infection. We present three initial analyses of these measurements: an overview of the CSF protein features of the sample; correlations of the CSF proteins with CSF HIV-1 RNA and neurofilament light chain protein (NfL) concentrations; and comparison of the CSF proteins in HIV-associated dementia ( HAD ) and neurosymptomatic CSF escape ( NSE ). These reveal a complex but coherent picture of CSF protein changes that includes highest concentrations of many proteins during CNS injury in the HAD and NSE groups and variable protein changes across the course of neuroasymptomatic systemic HIV-1 progression, including two common patterns, designated as lymphoid and myeloid patterns, related to the principal involvement of their underlying inflammatory cell lineages. Antiretroviral therapy reduced CSF protein perturbations, though not always to control levels. The dataset of these CSF protein measurements, along with background clinical information, is posted online. Extended studies of this unique dataset will provide more detailed characterization of the dynamic impact of HIV-1 infection on the CSF proteome across the spectrum of HIV-1 infection, and further the mechanistic understanding of HIV-1-related CNS pathobiology.

Effect of high irradiance and short exposure times on the depth of cure of six bulk-fill resin composites.

This study examined the effect of high irradiance and short exposure times on the depth of cure of six resin-based composites (RBCs). Bluephase PowerCure and the Valo X light-curing units (LCUs) were used to photocure bulk-fill RBCs for their recommended exposure times: Admira Fusion x-tra (AFX/20s), Aura Bulk Fill (ABF/20s), Filtek One Bulk Fill (FOB/20s), Opus Bulk Fill APS (OBF/30s), Tetric EvoCeram Bulk Fill (TEC/10s) and Tetric PowerFill (TPF/10s). In addition, all bulk-fill RBCs were tested for depth of cure with one short 3 s exposure time from the Bluephase PowerCure or the Valo X in the Xtra Power mode. The RBCs (n = 10 per RBC) were inserted into a 4 mm diameter metal mold and covered by a polyester strip before being photocured. After 24 h of storage, uncured RBC was scraped away to determine the depth of cure of the RBCs. None of the RBCs achieved a 4 mm depth of cure. The depth of cure of TEC and TPF was unaffected by the exposure times (recommended or short) when using the Valo X. The depth of cure of AFX/20s, AFX/Xtra Power, ABF/Xtra Power, FOB/Xtra Power, and OBF/30s RBCs was greater when using Valo X compared to the Bluephase PowerCure. It was concluded that short exposure times can reduce depth of cure and should only be used for some RBCs.

Exploring the dynamics of adult Axin2 cell lineage integration into dentate gyrus granule neurons.

The Wnt pathway plays critical roles in neurogenesis. The expression of Axin2 is induced by Wnt/ß-catenin signaling, making this gene a reliable indicator of canonical Wnt activity. We employed pulse-chase genetic lineage tracing with the Axin2-CreERT2 allele to follow the fate of Axin2+ lineage in the adult hippocampal formation. We found Axin2 expressed in astrocytes, neurons and endothelial cells, as well as in the choroid plexus epithelia. Simultaneously with the induction of Axin2 fate mapping by tamoxifen, we marked the dividing cells with 5-ethynyl-2'-deoxyuridine (EdU). Tamoxifen induction led to a significant increase in labeled dentate gyrus granule cells three months later. However, none of these neurons showed any EdU signal. Conversely, six months after the pulse-chase labeling with tamoxifen/EdU, we identified granule neurons that were positive for both EdU and tdTomato lineage tracer in each animal. Our data indicates that Axin2 is expressed at multiple stages of adult granule neuron differentiation. Furthermore, these findings suggest that the integration process of adult-born neurons from specific cell lineages may require more time than previously thought.

Focused Ultrasound Blood-Brain Barrier Opening Arrests the Growth and Formation of Cerebral Cavernous Malformations.

BACKGROUND: Cerebral cavernous malformations (CCM) are vascular lesions within the central nervous system, consisting of dilated and hemorrhage-prone capillaries. CCMs can cause debilitating neurological symptoms, and surgical excision or stereotactic radiosurgery are the only current treatment options. Meanwhile, transient blood-brain barrier opening (BBBO) with focused ultrasound (FUS) and microbubbles is now understood to exert potentially beneficial bioeffects, such as stimulation of neurogenesis and clearance of amyloid-ß. Here, we tested whether FUS BBBO could be deployed therapeutically to control CCM formation and progression in a clinically-representative murine model. METHODS: CCMs were induced in mice by postnatal, endothelial-specific Krit1 ablation. FUS was applied for BBBO with fixed peak-negative pressures (PNPs; 0.2-0.6 MPa) or passive cavitation detection-modulated PNPs. Magnetic resonance imaging (MRI) was used to target FUS treatments, evaluate safety, and measure longitudinal changes in CCM growth after BBBO. RESULTS: FUS BBBO elicited gadolinium accumulation primarily at the perilesional boundaries of CCMs, rather than lesion cores. Passive cavitation detection and gadolinium contrast enhancement were comparable in CCM and wild-type mice, indicating that Krit1 ablation does not confer differential sensitivity to FUS BBBO. Acutely, CCMs exposed to FUS BBBO remained structurally stable, with no signs of hemorrhage. Longitudinal MRI revealed that FUS BBBO halted the growth of 94% of CCMs treated in the study. At 1 month, FUS BBBO-treated lesions lost, on average, 9% of their pre-sonication volume. In contrast, non-sonicated control lesions grew to 670% of their initial volume. Lesion control with FUS BBBO was accompanied by a marked reduction in the area and mesenchymal appearance of Krit mutant endothelium. Strikingly, in mice receiving multiple BBBO treatments with fixed PNPs, de novo CCM formation was significantly reduced by 81%. Mock treatment plans on MRIs of patients with surgically inaccessible lesions revealed their lesions are amenable to FUS BBBO with current clinical technology. CONCLUSIONS: Our results establish FUS BBBO as a novel, non-invasive modality that can safely arrest murine CCM growth and prevent their de novo formation. As an incisionless, MR image-guided therapy with the ability to target eloquent brain locations, FUS BBBO offers an unparalleled potential to revolutionize the therapeutic experience and enhance the accessibility of treatments for CCM patients.

Focused ultrasound ablation of melanoma with boiling histotripsy yields abscopal tumor control and antigen-dependent dendritic cell activation.

Background: Boiling histotripsy (BH), a mechanical focused ultrasound ablation strategy, can elicit intriguing signatures of anti-tumor immunity. However, the influence of BH on dendritic cell function is unknown, compromising our ability to optimally combine BH with immunotherapies to control metastatic disease. Methods: BH was applied using a sparse scan (1 mm spacing between sonications) protocol to B16F10-ZsGreen melanoma in bilateral and unilateral settings. Ipsilateral and contralateral tumor growth was measured. Flow cytometry was used to track ZsGreen antigen and assess how BH drives dendritic cell behavior. Results: BH monotherapy elicited ipsilateral and abscopal tumor control in this highly aggressive model. Tumor antigen presence in immune cells in the tumor-draining lymph nodes (TDLNs) was ~3-fold greater at 24h after BH, but this abated by 96h. B cells, macrophages, monocytes, granulocytes, and both conventional dendritic cell subsets (i.e. cDC1s and cDC2s) acquired markedly more antigen with BH. BH drove activation of both cDC subsets, with activation being dependent upon tumor antigen acquisition. Our data also suggest that BH-liberated tumor antigen is complexed with damage-associated molecular patterns (DAMPs) and that cDCs do not traffic to the TDLN with antigen. Rather, they acquire antigen as it flows through afferent lymph vessels into the TDLN. Conclusion: When applied with a sparse scan protocol, BH monotherapy elicits abscopal melanoma control and shapes dendritic cell function through several previously unappreciated mechanisms. These results offer new insight into how to best combine BH with immunotherapies for the treatment of metastatic melanoma.

Thermography and conversion of fast-cure composite photocured with quad-wave and laser curing lights compared to a conventional curing light.

OBJECTIVES: This study investigated effects of the different emittance-mode protocols from three light curing units (LCUs): (i) a Laser (Monet); (ii) a quad-wave (PinkWave); (iii) a conventional LED (Elipar S10) on the temperature rise (ΔT) and degree of conversion (DC) when photo-curing fast or conventional bulk-fill resin-based composites (RBC). The aim was to correlate ΔT and DC, and the radiant exposure delivered to RBC specimens. METHODS: A 3D-printed resin mold of 4 mm depth was filled with two bulk-fill RBCs: Tetric PowerFill® (fast photo-polymerised composite) (TPF) or Tetric EvoCeram® Bulk-Fill (EVO). Three LCUs were used: (i) Monet laser for 1 s and 3 s (MONET-1 s, MONET-3 s); (ii) PinkWave quad-wave used for 3 s in Boost mode (PW-3 s) and for 20 s in standard mode (PW-20 s); (iii) Elipar S10 for 5 s (S10-5 s) and for 20 s in standard mode (S10-20 s). 2-dimensional temperature maps were obtained before, during and for 60 s after the LCU had turned off using a thermal imaging camera. Thermal changes were analysed at five depths: (0, 1, 2, 3, and 4 mm from the top surface of the RBC). The maximum temperature rise (Tmax) and the mean temperature rise (ΔT) were determined. Cylindrical-shaped specimens were prepared from each material using a stainless-steel split mold (4 × 4 mm) and light-cured with the same protocols. The DC was measured for 120 s and at 1 h after LCU had turned off using Fourier Transform Infrared Spectroscopy (FTIR). Data were analysed using Three-way ANOVA, One-way ANOVA, independent t-tests, and Tukey post-hoc tests (p < 0.05). RESULTS: Radiant exposures delivered by the various irradiation protocols were between 4.5-30.3 J/cm2. Short exposure times from MONET-1 s and PW-3 s delivered the lowest radiant exposures (4.5 and 5.2 J/cm2, respectively) and produced the lowest ΔT and DC. The longer exposure times in the standard modes of PW-20 s, S10-20 s, and MONET-3 s produced the highest Tmax, ΔT, and DC for both composites. The ΔT range among composites at different depths varied significantly (31.7-49.9 °C). DC of TPF ranged between 30-65% and in EVO between 15.3-56%. TPF had higher Tmax, ΔT for all depths and DC compared to EVO, across the LCU protocols (p < 0.05), except for PW-20 s and MONET-3 s. The coronal part of the restorations (1-2 mm) had the highest ΔT. There was a positive correlation between ΔT and DC at 4-mm depth after 120 s SIGNIFICANCE: Longer, or standard, exposure times of the LCUs delivered greater radiant exposures and had higher DC and ΔT compared to shorter or high-irradiance protocols. The fast photo-polymerised RBC had comparatively superior thermal and conversion outcomes when it received a high irradiance for a short time (1-5 s) compared to the conventional Bulk-Fill RBC.

Response to comment on 'The clinical pharmacology of tafenoquine in the radical cure of Plasmodium vivax malaria: An individual patient data meta-analysis'.

In our recent paper on the clinical pharmacology of tafenoquine (Watson et al., 2022), we used all available individual patient pharmacometric data from the tafenoquine pre-registration clinical efficacy trials to characterise the determinants of anti-relapse efficacy in tropical vivax malaria. We concluded that the currently recommended dose of tafenoquine (300 mg in adults, average dose of 5 mg/kg) is insufficient for cure in all adults, and a 50% increase to 450 mg (7.5 mg/kg) would halve the risk of vivax recurrence by four months. We recommended that clinical trials of higher doses should be carried out to assess their safety and tolerability. Sharma and colleagues at the pharmaceutical company GSK defend the currently recommended adult dose of 300 mg as the optimum balance between radical curative efficacy and haemolytic toxicity (Sharma et al., 2024). We contend that the relative haemolytic risks of the 300 mg and 450 mg doses have not been sufficiently well characterised to justify this opinion. In contrast, we provided evidence that the currently recommended 300 mg dose results in sub-maximal efficacy, and that prospective clinical trials of higher doses are warranted to assess their risks and benefits.

Clinical Value of Timely Targeted Therapy for Patients With Advanced Non-Small Cell Lung Cancer With Actionable Driver Oncogenes.

BACKGROUND: A recent real-world study observed that 24% of patients with advanced non-small cell lung cancer (aNSCLC) with actionable driver oncogenes (ADOs) initiated nontargeted therapies before biomarker test results became available. This study assessed the clinical impact of the timing of first-line (1L) targeted therapies (TTs) in aNSCLC. MATERIALS AND METHODS: This retrospective analysis of a nationwide electronic health record-derived deidentified database included patients aged ≥18 years diagnosed with aNSCLC with ADOs (ALK, BRAF, EGFR, RET, MET, ROS-1, and NTRK) from January 1, 2015, to October 18, 2022, by biomarker testing within 90 days after advanced diagnosis and received 1L treatment. Cohorts were defined by treatment patterns ≤42 days after test results: "Upfront TT" received 1L TT ≤42 days; "Switchers" initiated 1L non-TT before or after testing but switched to TT ≤42 days; and "Non-switchers" initiated non-TT before or after testing and did not switch at any time. Adjusted multivariate Cox regression evaluated real-world progression-free survival, real-world time to next treatment or death, and real-world overall survival. RESULTS: A total of 3540 patients met the study criteria; 78% were treated in a community setting, and 50% underwent next-generation sequencing (NGS). There was no significant difference in outcomes between Switchers and Upfront TT; inferior outcomes were observed in Non-switchers versus Upfront TT. CONCLUSION: Our findings demonstrated improved outcomes with upfront 1L TT versus non-TT in patients with aNSCLC with ADOs and observed timely switching to TT after biomarker test result had similar outcomes to Upfront TT. Opportunities remain to improve the use of NGS for early ADO identification and determination of 1L TT.