Genentech

Basic fibroblast growth factor (bFGF) and Transforming growth factor-beta (TGF-β) are key regulators of human pluripotent stem cell (hPSC) maintenance, supporting pluripotency and self-renewal. bFGF is particularly critical for sustaining the undifferentiated state and is commonly supplied through feeder-derived conditioned media. Similarly, TGF-β promotes hPSC expansion by modulating signaling pathways and contributing to a supportive stem cell niche. In this study, we investigated how the quality and variability of these growth factors influence hPSC culture performance. To address this, we developed and applied multiple physicochemical characterization methods-including size exclusion and reverse-phase chromatography-to assess growth factor purity and identify impurities across different material sources. Our findings show that certain post-translational modifications in TGF-β (e.g., oxidized variants) did not measurably affect hPSC culture. However, high temperature-dependent instability of bFGF preparations significantly altered hPSC morphology and growth. These findings underscore the need for improved quality control of growth factor components in culture media to ensure consistent hPSC maintenance, thus decreasing variability across experiments. This study highlights the value of correlating analytical physicochemical data with process performance, thereby advancing material understanding, enabling more efficient process development, and facilitating the identification of critical material attributes that affect the quality of cell therapy products.

The accelerated approval (AA) pathway expedites treatment access for patients with unmet medical needs. However, evidence on the program’s impact is lacking. This study assessed real-world clinical outcomes for AA drugs in solid tumor cancers. A nationwide electronic health record (EHR)-derived deidentified database was utilized to explore the impact of the AA pathway in patients with selected solid tumors from 2011 to 2023. Included patients received AA or non-AA products with the same indication. Outcomes were real-world progression-free survival (rwPFS) and overall survival (OS). Population-level health impacts were estimated via market share and real-world progression-free life years (rwPFLYs) and life years (LYs). Twenty-three AA indications were studied. At study completion, 83% of indications had their clinical benefit verified, whereas 17% had been withdrawn from the market. Overall, 78% of indications demonstrated significant improvements in rwPFS or OS, whereas none reported significantly worse rwPFS versus controls. In total, 44% and 48% of indications had >30% rwPFS and OS improvement, respectively. More indications with sample sizes >100 in the AA cohort demonstrated significant rwPFS gains [80% (12/15)] versus cohorts <100 [38% (3/8)]. An estimated 118,107 patients received AA drugs during their AA window, resulting in 56,399 rwPFLYs and 50,848 LYs gained. Most solid tumor AA indications were associated with improved clinical outcomes, providing substantial US population-level health gains. This EHR analysis provides insight into whether the AA pathway appropriately targets promising beneficial therapies. Expanding beyond randomized controlled trial evidence and conventional primary endpoints may be necessary to verify clinical benefit, particularly in rare tumors.Significance:This study (the first examining the real-world outcomes associated with the AA period) found improved clinical outcomes for most AA drugs studied compared with existing treatments in solid tumor cancers. The improvements observed suggest potential health improvements at the US population level and the possibility of the need to look beyond randomized controlled evidence and traditional primary endpoints to verify the clinical benefit of AA drugs for relatively rare conditions.

SummaryExisting cancer-associated variant databases contain biases arising from duplicate entries and the inclusion of targeted sequencing panels, which interfere with accurate estimation somatic mutation frequency in cancer cohorts. To address this, we developed the Database of Recurrent Mutations (DORM), a web resource derived exclusively from whole-genome and whole-exome sequencing data. By filtering out targeted screens and non-recurrent variants, our analysis reveals that mutation recurrence significantly correlates with oncogenic activity, loss of tumor suppressor function, and unfavorable patient prognosis. In a pan-cancer analysis of EGFR, DORM identified frequent mutations outside the kinase domain that are underrepresented in other databases. This resource offers a streamlined, unbiased platform for mutation frequency analysis, enhancing biomarker discovery and the assessment of clinical variant significance.

FoxQ2 is a highly conserved Forkhead-box transcription factor expressed anteriorly in cnidarians and bilaterians, yet its evolution is marked by rapid divergence and lineage-specific duplications or losses. Moreover, its presence and localization in vertebrate groups remains unclear. To reconcile these conflicting reports of conservation and divergence, we combine phylogenetic and synteny analyses of FoxQ2 sequences from 21 animal phyla. We uncover three ancient FoxQ2 paralogs in bilaterians—FoxQ2I, FoxQ2II, and FoxQ2III. All three were present in the chordate ancestor, and two are retained in vertebrates, indicating a richer FoxQ2 repertoire in vertebrates than previously recognized. To assess FoxQ2 expression, we analyzed mollusk, acoel, amphioxus, and zebrafish single-cell transcriptomic datasets, and conducted fluorescent in situ hybridization in amphioxus, lamprey, skate, zebrafish, and chicken. FoxQ2I and FoxQ2II show conserved anterior expression, while FoxQ2III is expressed in the gut endoderm in chordates, including amphioxus, lamprey, and skate. We also predict conserved transcription factor binding sites across amphioxus genera, revealing stage- and cell-type-specific regulatory interactions for FoxQ2I in deuterostomes. Overall, this work clarifies FoxQ2’s evolutionary history, identifies the endodermally expressed paralog FoxQ2III, and proposes that early duplication of FoxQ2I/II enabled subfunctionalization, driving the fast evolutionary rate of FoxQ2 sequences observed in bilaterians.

Multiplex immunofluorescence and immunohistochemistry (mIF/IHC) are increasingly employed antibody-based technologies that use tissue sparingly and facilitate the detection of co-localized or neighboring biomarkers. Specifically, these platforms enable spatial analyses of the tumor microenvironment as well as extended applications, for example, describing normal tissue anatomy, autoimmunity, infectious diseases, etc. mIF/IHC has greatly enhanced biomarker discovery efforts, and a growing number of studies suggest superiority to traditional IHC. Standardization of staining approaches, reporting of image analysis strategies and resultant data is critical for facilitating cross-study comparisons, validation, deployment, and generalization of findings. To address this challenge, The Society for Immunotherapy of Cancer (SITC) previously published two articles providing best practice guidelines for mIF/IHC staining, image analysis, and data sharing. Here, SITC convened stakeholders to develop the third article in the series, a consensus checklist for scientific reporting of mIF/IHC data to support and complement the best practice guidelines. The checklist includes critical components of mIF/IHC applications to be defined within publications such as detailed descriptions of analytical validation; image acquisition, selection, and registration methods; and cell clustering and spatial analysis strategies, amongst others. Such information will help with data reproducibility and comparison across studies towards future drug and assay development.

Radiolabeled human mass balance studies are crucial for identifying circulating metabolites and understanding drug absorption, excretion, and clearance pathways. Metabolite profiling involves quantifying all drug-related entities, including parent drug and metabolites in plasma and excreta, using extended liquid chromatography methods coupled with detection through scintillation counting, accelerator mass spectrometry, or non-radiolabeled approaches. Given the labor-intensive nature of sample extraction and analysis, we propose a new paradigm that maximizes gathering information through sample pooling strategies. Our proposal introduces sample pooling strategies by integrating both individual and pooled sample schemes, simplifying decisions, and consolidating existing knowledge into a cohesive document. This aligns with the low statistical power typically associated with mass balance studies that dose six to eight subjects. In metabolite profiling, it is common practice to pool samples either from the limited number of subjects participating in a human mass balance study or from different time points of sample collection. This approach improves efficiency while preserving data integrity. Pooling reduces resource constraints and enables the concentration of samples with relatively low radioactivity levels, resulting in higher quality metabolite profiles. Nevertheless, there are situations when analyzing samples from individual subjects or time points may be preferred. This proposal presents guidance and decision trees designed to facilitate informed decisions about sample pooling to maximize data quality of metabolite profiling in human mass balance studies while efficiently managing resources. These recommendations stem from discussions within the mass balance working group of the IQ Consortium.

The endoplasmic-reticulum (ER) transmembrane protein IRE1 mitigates ER stress through kinase-endoribonuclease and scaffolding activities. Cancer cells often co-opt IRE1 to facilitate growth. An IRE1-RNase inhibitor has entered clinical trials; however, recent work uncovered a significant nonenzymatic IRE1 dependency in cancer. To fully disrupt IRE1, we describe a proteolysis-targeting chimera (G6374) that couples an IRE1-kinase ligand to a compound that binds the ubiquitin Cullin-RING Ligase (CRL) substrate receptor, VHL. G6374 induces a stable, cooperative interaction between IRE1 and VHL, driving K48-linked ubiquitination on two principal lysine residues in the IRE1-kinase domain and inducing proteasomal IRE1 degradation. Cryogenic electron microscopy and mutagenesis studies reveal a 2:2 IRE1:VHL ternary-complex topology and critical interactional features, informing future designs. G6374 blocks growth of IRE1-dependent cancer cells irrespective of their dependency mode, while sparing IRE1-independent cells. We provide a proof-of-concept for VHL-based degradation of an ER-transmembrane protein, advancing strategies to fully disrupt IRE1.

IntroductionThe Inflation Reduction Act (IRA) authorizes Medicare price negotiation but includes a “special rule” deferring negotiation for biologics with “imminent” biosimilar competition. This study examined the potential impact of this provision on Medicare spending.MethodsWe modeled Medicare savings under three hypothetical scenarios. In Scenario 1, we applied historical price reductions following biosimilar entry (2017-2024) for 10 reference products and 30 biosimilars to forecast savings for ustekinumab (Stelara) after biosimilar entry. Scenario 2 estimated savings from the IRA's negotiated maximum fair price for ustekinumab. Scenario 3 modeled a modified IRA implementation policy in which ustekinumab was excluded from negotiation and replaced by palbociclib (Ibrance), an eligible high spend drug that was not selected for the first round of Medicare price negotiation.ResultsAcross all biologics in our sample, historic market-weighted prices declined to 40.3% of pre-entry levels within five years of biosimilar launch. Negotiating ustekinumab under the IRA yielded greater first-year savings, but cumulative savings were highest in the modified scenario-combining negotiated discounts for palbociclib with biosimilar-driven price declines for ustekinumab.ConclusionSelecting biologics with near-term biosimilar competition for IRA negotiation may produce short-term savings but forgo greater long-term savings achievable through competition.