Milk oligosaccharides (MOs) are essential for the development of mammalian offspring, yet their fine-scale structural evolutionary divergence remains unelucidated, largely due to isomeric complexity and the limitations o...Milk oligosaccharides (MOs) are essential for the development of mammalian offspring, yet their fine-scale structural evolutionary divergence remains unelucidated, largely due to isomeric complexity and the limitations of analytical methods. Here, we present GlycoBond X, an online platform that couples high-resolution separation with parallel structural characterization of glycan isomers through multi-stage chemical derivatization and RP-HPLC-MS/MS. By applying this strategy to four evolutionarily distinct mammals, we uncovered a conserved transition from acidic MO dominance (>75% in mouse and tree shrew) to predominantly fucosylated neutral MOs (>62% in macaque and human). GlycoBond X unveiled unprecedented structural diversity, including 22 unique fucosylation motifs and 66 previously undescribed human MOs. Notably, tree shrew MOs exhibited human-like structures and shared over 69% FUT2 sequence homology with primates. This study established a high-throughput, high-sensitivity platform, elucidated the adaptive structural evolution of oligosaccharides via evolutionary glycomics, and provided a foundation for exploring their biosynthetic pathways.
STING bridges innate and adaptive immunity to exert potent antitumor effects, and its agonists play emerging roles in combination therapies with tyrosine kinase inhibitors (TKIs). However, the interaction between STING a...STING bridges innate and adaptive immunity to exert potent antitumor effects, and its agonists play emerging roles in combination therapies with tyrosine kinase inhibitors (TKIs). However, the interaction between STING and receptor tyrosine kinases (RTKs) remains incompletely understood. Here, we identify VEGFR2 as a negative regulator of cGAMP-STING signaling. Upon cGAMP stimulation, both STING and VEGFR2 are activated. Activated VEGFR2 recruits and activates AKT1 to attenuate STING activation. Conversely, STING suppresses VEGFR2 phosphorylation, establishing a reciprocal inhibitory feedback loop. A cell-based screen reveals that the VEGFR2 inhibitor Ki8751 not only relieves VEGFR2-AKT1-mediated suppression but also activates MyD88-dependent NF-κB signaling to further amplify STING responses. Additionally, we show that Ki8751 synergizes with cGAMP to elicit robust, STING-dependent antitumor immunity in vivo. Our findings identify the VEGFR2-STING regulatory axis and provide a mechanistic rationale for co-targeting VEGFR2 and STING to improve cancer immunotherapy.
Chronic stress can induce pain and drive development of aversion, although the specific neural circuits that contribute to encoding such behavioral responses remain unclear. Here, we find that noradrenergic neurons in lo...Chronic stress can induce pain and drive development of aversion, although the specific neural circuits that contribute to encoding such behavioral responses remain unclear. Here, we find that noradrenergic neurons in locus coeruleus (NE) regulate the formation of allodynia and aversion under chronic restraint stress in mice, driven by enhanced activity of NE-projecting GABAergic neurons in the central amygdala (CeA). Selectively inhibiting this GABAergic CeA→LC pathway significantly alleviates pain sensitization and aversion behaviors, but without impacting chronic stress-associated depression-like behaviors. Additionally, these NE neurons receiving inputs from CeA monosynaptically activate glutamatergic neurons in the lateral periaqueductal gray (Glu) to mediate pain sensitization and aversion. This study thus defines a specific CeA→LC→LPAG pathway through which chronic stress leads to pain and aversion, without affecting depression-like behaviors, expanding our fundamental understanding of the circuit basis for stress-related pain.
The 5-methylcytosine (m5C) modification of mRNA mediates diverse cellular and viral functions. Epstein-Barr virus (EBV) is etiologically linked to multiple malignancies, including nasopharyngeal carcinoma (NPC), in which...The 5-methylcytosine (m5C) modification of mRNA mediates diverse cellular and viral functions. Epstein-Barr virus (EBV) is etiologically linked to multiple malignancies, including nasopharyngeal carcinoma (NPC), in which both viral latency and lytic replication play critical pathogenic roles. Here, we show that the transcripts derived from the EBV gene BHLF1 exhibit abundant m5C modification in patient-derived xenograft tissue as well as in NPC cells during the lytic stage of EBV infection. Mechanistically, NSUN2 mediates the m5C modification of EBV BHLF1 RNA, which not only enhances transcript stability but also promotes DNA methylation at the EBV origin of lytic replication (oriLyt) region near the m5C site through recruitment of DNA methyltransferase 1 (DNMT1) to facilitate BZLF1 binding, thereby promoting efficient EBV lytic replication. Taken together, our findings provide mechanistic insights into the roles of m5C modification in the EBV life cycle and highlight the importance of crosstalk between RNA m5C and DNA methylation in regulating viral replication.
Human cognition depends on the ability to flexibly recombine existing knowledge in new ways. Although this capacity for compositionality has traditionally been attributed to cortical networks, its broader neural basis re...Human cognition depends on the ability to flexibly recombine existing knowledge in new ways. Although this capacity for compositionality has traditionally been attributed to cortical networks, its broader neural basis remains unclear. We combine dimensionality reduction of task-based fMRI with recurrent neural network modeling to dissociate two processes underlying compositional cognition: specialized components; and the more general process of recombination. Across 87 participants performing a well-established compositional task, domain-specific cortical and anterior cerebellar regions support component processes, whereas a low-dimensional, highly integrated cortico-cerebellar network engages recombination processes consistently across diverse contexts. Recurrent neural networks trained to perform multiple cognitive tasks present similar functional signatures to the fMRI data, suggesting that low-dimensional recombination is a general solution for flexible compositional cognition. Our findings revise existing models of compositional cognition, highlighting interactions between component and recombination processes with cortico-cerebellar interactions serving as a mechanism for flexible, integrative task generalization.
Waters AM, Khatib TO, Papke B
… +34 more, Goodwin CM, Hobbs GA, Diehl JN, Yang R, Edwards AC, Walsh KH, Sulahian R, McFarland JM, Kapner KS, Gilbert TSK, Stalnecker CA, Javaid S, Barkovskaya A, Grover KR, Hibshman PS, Blake DR, Schaefer A, Nowak KM, Klomp JE, Hayes TK, Kassner M, Tang N, Tanaseichuk O, Chen K, Zhou Y, Kalkat M, Herring LE, Graves LM, Penn LZ, Yin HH, Aguirre AJ, Hahn WC, Cox AD, Der CJ
The etiology of Alzheimer's disease (AD) remains unclear but is likely driven by gene-environment interactions. We present a multi-organ untargeted metabolomics atlas (n = 2,271) paired with metagenomics data (n = 666) f...The etiology of Alzheimer's disease (AD) remains unclear but is likely driven by gene-environment interactions. We present a multi-organ untargeted metabolomics atlas (n = 2,271) paired with metagenomics data (n = 666) from two AD transgenic mouse models (3xTg and 5xFAD) under colonized and germ-free conditions. Systems-level analyses revealed clusters of dysregulated molecules across tissues, including carnitines, bile acids, B vitamins, neurotransmitters, and N-acyl lipids. Metabolic shifts were associated with the depletion of Akkermansia muciniphila and enrichment of Mucispirillum schaedleri in the 3xTg model. We identify previously unexplored carnitines linked to microbial metabolism of phenylalanine. Using tissueMASST-a mass spectrometry search tool we developed to translate animal-model findings into a human clinical context-we trace phenylacetyl-carnitine in human plasma and serum samples (n = 1,470) from independent cohorts, revealing associations with aging, cognitive impairment, and diminished memory performance. This public resource and associated tools will aid future research in AD etiology.
Acute myeloid leukemia is often associated with constitutive activation of the Src-family kinases, Hck, Lyn, and Fgr. Their modular SH3 and SH2 domains regulate kinase activity and signal transduction. Here, we show that...Acute myeloid leukemia is often associated with constitutive activation of the Src-family kinases, Hck, Lyn, and Fgr. Their modular SH3 and SH2 domains regulate kinase activity and signal transduction. Here, we show that regulatory domain dynamics critically influence both inhibitor sensitivity and leukemogenic signaling. We modified the Fgr SH2-kinase linker to enhance intramolecular SH3 engagement, shifting the conformational ensemble to the closed state. This shift increased the K for ATP and enhanced the potency of ATP-site inhibitors in vitro. Human myeloid cells expressing these constrained Fgr variants exhibited heightened sensitivity to ATP-site inhibitors in terms of growth arrest. These cells also demonstrated impaired bone marrow engraftment in vivo, suggesting a key role for Fgr dynamics and SH3-dependent signaling in leukemia cell survival within this niche. Small molecules that similarly restrict Src-family kinase regulatory domain dynamics may provide a new therapeutic approach to AML and other cancers linked to these kinases.
Alyahyay M, Ammer JJ, Kalweit G
… +12 more, Zhu H, Adzemovic A, Kalweit M, Karvat G, Schneider A, Tong Y, Döbrössy MD, Jäckel Z, Akmese C, Vlachos A, Boedecker J, Diester I
Preparing and initiating movements at the right time is critical for goal-directed behavior. Before movement execution, motor cortical areas exhibit preparatory activity, which decreases from premotor to primary motor ar...Preparing and initiating movements at the right time is critical for goal-directed behavior. Before movement execution, motor cortical areas exhibit preparatory activity, which decreases from premotor to primary motor areas. During the shift from preparation to execution activity in neural state space transitions from movement-null to movement-potent dimensions. However, the circuit-level mechanisms underlying this shift remain unresolved. Here, we demonstrate that projections from the rat premotor cortex (rostral forelimb area [RFA]) to the primary motor cortex (caudal forelimb area [CFA]) encode primarily pre-movement activity. Optogenetic inhibition of these projections has behavioral effects comparable to inhibiting either RFA or CFA alone. During preparation, RFA projections enhance and suppress CFA neurons similarly, affecting activity along CFA's preparatory dimension. During movement, RFA's influence shifts predominantly to excitatory, aligning with CFA's movement-potent dimension. These results establish a mechanistic link between neural state space concepts and underlying circuit mechanisms, providing an intuitive model for movement control.
Microbiota-derived metabolites are central mediators between commensal microbes and host immune system at mucosal barrier surfaces. Insights from mouse models have revealed precise molecular mechanisms by which numerous...Microbiota-derived metabolites are central mediators between commensal microbes and host immune system at mucosal barrier surfaces. Insights from mouse models have revealed precise molecular mechanisms by which numerous metabolites, including short-chain fatty acids, tryptophan catabolites and bile acid derivatives, regulate epithelial integrity, innate immune tone, and adaptive immunity and tolerance. Parallel studies in humans increasingly confirm these pathways and link metabolite dysregulation to diseases, such as inflammatory bowel disease, asthma, and atopic dermatitis. This review synthesizes current understanding of how microbial metabolites orchestrate gastrointestinal barrier immunity, while also integrating emerging insights into the gut-lung and gut-skin axes. Crucially, we examine these interactions through a developmental lens, highlighting how metabolite exposure during critical early-life "windows of opportunity" shapes long-term immune trajectories. We integrate evidence from experimental models and human data to highlight conserved mechanisms, species-specific divergences, and the therapeutic potential of targeting these metabolic pathways as strategies to promote barrier health and durable immune homeostasis.
Ghafouri E, Hill K, Mellor N
… +24 more, Leftley N, Sato EM, Banda J, Lale A, Bansod A, Wilson M, Pandey BK, Truskina J, Oh J, Han J, Kilic A, Wells D, Péret B, Swarup R, Hagen G, Guilfoyle T, Vissenberg K, Berleth T, Vernoux T, Bishopp A, Fukaki H, Bennett MJ, Goh T, Bhosale RA
Root gravitropism is regulated by an auxin gradient, causing the root to reorient downwards with gravity. Transcriptional activator auxin response factors (ARFs), such as ARF5, ARF6, ARF7, ARF8, and ARF19, play key roles...Root gravitropism is regulated by an auxin gradient, causing the root to reorient downwards with gravity. Transcriptional activator auxin response factors (ARFs), such as ARF5, ARF6, ARF7, ARF8, and ARF19, play key roles in auxin signaling in seedling root tissues. ARF7 and ARF19 are both required for root gravitropism, but their regulatory relationship with each other and other activating ARFs remains unknown. Here, we identify that rapid and transient upregulation of ARF19 during gravitropic stimulus is functionally important. ARF7 and ARF8 are required to activate ARF19 upregulation via auxin response elements (AuxREs) in its promoter, while our data indicate that ARF19 does not self-activate. ARF7 can replace ARF19 but not vice versa. Although ARF7 shares overlapping targets with ARF19, they also have distinct functionally important targets, such as ARF19. Our results show that ARF7 and ARF19 have sub-functionalized, with ARF19 acting as an auxin response enhancer for a subset of ARF7 target genes.
Wang Y, Kuo HC, Kuang X
… +43 more, Yao S, Lesnar P, Ng L, Li Y, El-Hifnawi L, Chen N, Zhang K, Li EM, Ben-Simon Y, Ding SL, Wang Q, Karlsson T, Dalley R, Williams G, Xiong W, Chen C, Chen K, Huang Z, Yu ZF, Xu WJ, Ahmadinia L, Walling-Bell S, Andrade J, Gliko O, Mallory M, Farrel C, Sutton B, Jin K, Yao Z, Halterman C, Ronellenfitch K, Esposito L, Sunkin S, Kruse L, Luo Q, Gong H, Li A, Li X, Qu J, Choi H, Mihalas S, Zhuang J, Sorensen S
Neocortex contains diverse excitatory neurons whose dendritic and axonal architectures shape computation and long-range communication, yet a unified framework linking neuronal structure, molecular identity, and brain-wid...Neocortex contains diverse excitatory neurons whose dendritic and axonal architectures shape computation and long-range communication, yet a unified framework linking neuronal structure, molecular identity, and brain-wide connectivity is lacking. Here, we combine complete whole-neuron morphology (WNM) reconstructions with paired genetic identities across 15 mouse cortical areas to define organizing principles of cortical wiring at single-cell resolution. We identify 10 stable excitatory cell types with conserved morphology and predominant genetic correspondence. Their full axonal target spectra reveal four output architectures and cell-type-resolved principles of corticocortical and cortico-subcortical organization, including modular convergence in downstream targets, distinct spatial topographical rules across corticocortical and corticofugal pathways, and hierarchical organization better predicted by targeting probability and projection distribution than projection strength, while refining bulk-derived projectomes by resolving artifacts from bulk approaches. Together, these findings establish WNM as a principled axis for defining cortical cell types as building blocks of large-scale connectomes.
Breen MS, Tao R, Yang A
… +10 more, Wang X, Amini P, Rodriguez de Los Santos M, Brandtjen AC, Deep-Soboslay A, Kaye WH, Hyde TM, Kleinman JE, Buxbaum JD, Grice DE
Eating disorder (ED) and obsessive-compulsive disorder (OCD) exhibit clinical and genetic overlap, yet whether they converge at the molecular level in the human brain is unknown. We perform large-scale transcriptomic pro...Eating disorder (ED) and obsessive-compulsive disorder (OCD) exhibit clinical and genetic overlap, yet whether they converge at the molecular level in the human brain is unknown. We perform large-scale transcriptomic profiling of the dorsolateral prefrontal cortex (DLPFC) and caudate in postmortem tissue from 86 controls, 57 individuals with ED, and 27 with OCD. ED shows robust, region-specific transcriptional dysregulation (102 differentially expressed genes [DEGs] in DLPFC and 222 in caudate at FDR <1%) that replicates in an independent cohort. OCD shows no single-cohort DEGs, but meta-analysis across three datasets identifies 57 caudate-associated genes. Despite these differences, transcriptome-wide effects strongly correlate between ED and OCD (DLPFC r = 0.67; caudate r = 0.75), indicating shared molecular pathology. Joint ED + OCD analysis identifies 233 DEGs in DLPFC and 815 in caudate, implicating GABAergic signaling, neuroendocrine regulation, mitochondrial metabolism, and CHD8-associated networks. Genetically regulated expression analyses identifies five genes (WDR6, NCKIPSD, P4HTM, DALRD3, and SHISA5) with convergent risk associations across disorders and brain regions, all mapping to a gene-dense region on chromosome 3. These findings define a shared cortico-striatal transcriptional architecture and identify candidate genes for transdiagnostic intervention.
WLJP-025p, a homogeneous polysaccharide from Lonicera japonica Thunb., alleviates experimental ulcerative colitis in mice by reshaping the gut microbiota and restoring intestinal spermidine. Microbial fermentation, micro...WLJP-025p, a homogeneous polysaccharide from Lonicera japonica Thunb., alleviates experimental ulcerative colitis in mice by reshaping the gut microbiota and restoring intestinal spermidine. Microbial fermentation, microbiota depletion, and microbiota transplantation support a microbiota-dependent mechanism. Microbiota-derived spermidine interacts with HADHA and partially reverses inflammatory HADHA-associated metabolic reprogramming, as shown by restored fatty acid oxidation (FAO), reduced lactate accumulation, improved basal respiration and etomoxir-sensitive oxygen consumption rate (OCR), and preserved epithelial integrity in Caco-2 cells and intestinal organoids. In vivo, WLJP-025p dose-dependently reduces colitis pathology and inflammation, partially reverses FAO-related metabolic disruption, and promotes epithelial maturation and barrier integrity. Serum biochemical and protein analyses further support reduced systemic inflammation and improved mucosal homeostasis. These findings define a microbiota-derived spermidine-HADHA axis that supports mucosal homeostasis in ulcerative colitis.
Co-transcriptional splicing plays a crucial role in eukaryotic gene expression. However, its specific effects on growth, development, and stress responses in animals and plants are still not fully understood. In this stu...Co-transcriptional splicing plays a crucial role in eukaryotic gene expression. However, its specific effects on growth, development, and stress responses in animals and plants are still not fully understood. In this study, we screen for a pair of transcription factor and splicing factor interactions (AtCDC5 and AtSYF2) and find that AtCDC5 and AtSYF2, as splicing factors, can jointly regulate the co-transcriptional splicing of FLC, thereby affecting plant flowering time. Both proteins can bind to FLC pre-mRNA and participate in splicing while also influencing Pol II recruitment and R-loop formation. Additionally, the N terminus of AtCDC5 can bind to the promoter of FLC and act as a transcription factor to induce its expression. Mutant and complementary line analyses confirm the function of these factors as negative regulators of flowering time. These findings provide insights into the molecular mechanisms of co-transcriptional splicing in plant growth and development.
The assembly of cellular microtubules relies heavily on γ-tubulin ring complex (γTuRC), a macromolecular assembly of γ-tubulin and associated proteins that serves as a nucleation template. Here, we identify that within γ...The assembly of cellular microtubules relies heavily on γ-tubulin ring complex (γTuRC), a macromolecular assembly of γ-tubulin and associated proteins that serves as a nucleation template. Here, we identify that within γTuRC, γ-tubulin undergoes mitosis-specific phosphorylation at the conserved residue Ser364. This phosphorylation is mediated by Cdk1/cyclin B and occurs exclusively in cytoplasmic γTuRC, but not in γTuRC associated with mitotic spindles. Functionally, Ser364 phosphorylation strongly suppresses the microtubule-nucleating activity of γTuRC. Although γTuRC activity is essential for spindle microtubule assembly, disrupting Ser364 phosphorylation by expressing a non-phosphorylatable γ-tubulin mutant leads to defective spindle formation and chromosome segregation. Ser364 phosphorylation establishes spatial control over microtubule nucleation by inactivating cytoplasmic γTuRC, while spindle-associated γTuRC remains unphosphorylated and functionally active, consistent with the recently identified inhibitory control of spindle-localized Cdk1/cyclin B. This γTuRC regulation acts together with other Cdk1/cyclin B actions to eliminate non-spindle microtubules and support spindle assembly. Our findings reveal that Ser364 phosphorylation provides precise microtubule control for mitotic progression.
Gatesman TA, Varadharajan S, Johnson BJ
… +36 more, Duchatel RJ, Pathak R, Cruz AF, Wolfe HG, Halligan K, Garton LG, Johnson MA, Li F, Chilukuri A, Tjoa K, Halbert ME, Plute TJ, Jane EP, David Premkumar DR, Perez JL, Michel JJ, Gersey ZC, Shiva S, Kohanbash G, Vicha A, Zapotocky M, Felker J, Carlson S, Pearce TM, Gittes GK, Hu B, Prochownik EV, Wang X, Wu YL, Abel TJ, Mantica M, Pollack IF, Rich JN, Dun MD, Mack SC, Agnihotri S
MYCN functions as a developmental oncogene, but its role in pediatric high-grade gliomas (pHGGs) remains unclear. In co-operation with Trp53 and Pten loss, MYCN initiates tumorigenesis and establishes an origin for MYCN-...MYCN functions as a developmental oncogene, but its role in pediatric high-grade gliomas (pHGGs) remains unclear. In co-operation with Trp53 and Pten loss, MYCN initiates tumorigenesis and establishes an origin for MYCN-driven pHGGs. This transformation creates a vulnerability to PI3K and mTOR inhibition. However, prolonged treatment drives adaptive resistance through MYCN protein rebound, mediated by the attenuation of IGFBP5 and the induction of insulin-like growth factor 2. Although insulin pathway feedback has been implicated in resistance to PI3K targeted therapies, MYCN emerges as the central node of this adaptive program. Resistance can be overcame by sustained MYCN suppression using PI3K and mTOR inhibitors, combined with insulin-like growth factor 1 receptor and insulin receptor inhibitors or dietary intervention. A degradation-resistant MYCN isoform abolishes this response, establishing MYCN as both an initiating oncogene and a resistance driver and revealing a mechanistically defined therapeutic vulnerability.
Goodson BA, Montenegro Vazquez V, Doyle A
… +11 more, Kolaczkowski OM, Liu R, Jia J, Ishida M, Ye C, Kell AM, Bradfute SB, Rosas Lemus M, Wang H, Han X, Pu J
Cholesterol homeostasis depends on lysosomes liberating cholesterol from degraded lipids. We show that SARS-CoV-2 blocks lysosomal cholesterol egress through the viral protein ORF3a. ORF3a binds the HOPS subunit VPS39 vi...Cholesterol homeostasis depends on lysosomes liberating cholesterol from degraded lipids. We show that SARS-CoV-2 blocks lysosomal cholesterol egress through the viral protein ORF3a. ORF3a binds the HOPS subunit VPS39 via the W193 and Y184 residues. Disrupting this interface restores cholesterol trafficking. Mechanistically, the ORF3a-VPS39 interaction exerts dual effects. First, it traps the retromer complex on endolysosomes, preventing endosome-to-Golgi recycling and mislocalizing the cholesterol transporter NPC2. Retromer deletion reproduced these defects, whereas the ORF3a W193A mutant restored retromer trafficking. Second, ORF3a-VPS39 interaction reduces bis(monoacylglycerol)phosphates (BMPs), lysosomal lipids required for cholesterol egress, by disrupting the transfer of their precursor, phosphatidylglycerols, from mitochondria. Lipidomics revealed increased mitochondrial and decreased lysosomal phosphatidylglycerol metabolites. Disturbing autophagy or mitochondrion-derived vesicles did not alter BMP levels, whereas ORF3a reduced mitochondrion-lysosome membrane contacts. These findings identify dual functions of VPS39-regulating retromer trafficking and BMP biosynthesis-and also reveal how ORF3a blocks lysosomal cholesterol egress.
Eggs of many species accumulate thousands of dormant mRNAs that are translated after fertilization at specific times and locations to direct development. However, how embryos coordinate translation of these mRNAs remains...Eggs of many species accumulate thousands of dormant mRNAs that are translated after fertilization at specific times and locations to direct development. However, how embryos coordinate translation of these mRNAs remains unclear. In this study, we identify sequential waves of translation critical for proper development progression. The first wave occurs within 1 h and includes translation of ewsr1b mRNA that harbors a short 3' untranslated region (UTR) comprising 16 nucleotides. The resulting Ewsr1b protein triggers the second translation wave through binding cytoplasmic mRNAs, including pou5f3, which encodes a transcription factor promoting zygotic genome activation. In contrast, HuR and Syncrip repress translation until the first and second waves, respectively. ewsr1b mRNA that has a long 3' UTR is translated in the second wave, and the 3' UTR's length determines protein localization and function. Overall, our findings reveal previously unknown molecular principles that coordinate translation timings and protein functions to drive long-term, multilayered processes.
An efficacious HIV vaccine will need to generate broadly neutralizing antibodies (bnAbs) against distinct viral epitopes. To facilitate this, immunogens targeting precursor B cells of bnAbs have been developed. With this...An efficacious HIV vaccine will need to generate broadly neutralizing antibodies (bnAbs) against distinct viral epitopes. To facilitate this, immunogens targeting precursor B cells of bnAbs have been developed. With this strategy, individual immunogens can even target multiple lineages, thereby beneficially limiting the number of immunogens needed for a multi-bnAb-generating vaccine. However, it is unclear whether this approach diminishes the responses compared with isolated targeting of lineages with distinct immunogens. Here, we address this using an in silico model of naive B cell activation and affinity maturation in germinal centers. By incorporating the (1) precursor properties and (2) epitope masking by antibodies obtained from germinal center-derived plasma cells, the model recapitulated features of bnAb lineage evolution as seen in pre-clinical mouse models. Our model predicts that under physiologically relevant conditions, priming of multiple bnAb lineages with a single immunogen can be additive, thus having implications for further testing and development of multi-lineage-targeting immunogens.