Broad-spectrum Ebola vaccine
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arXiv:2502.07209v4 Announce Type: replace Abstract: Physics-Informed Neural Networks (PINNs) seek to solve partial differential equations (PDEs) with deep learning. Mainstream approaches that deploy fully-connected multi-layer deep learning architectures require prolonged training to achieve even moderate accuracy, while recent work on feature engineering allows higher accuracy and faster convergence. This paper introduces SAFE-NET, a Single-layered Adaptive Feature Engineering NETwork that achieves orders-of-magnitude lower errors with far fewer parameters than baseline feature engineering methods. SAFE-NET returns to basic ideas in machine learning, using Fourier features, a simplified single hidden layer network architecture, and an effective optimizer that improves the conditioning of the PINN optimization problem. Numerical results show that SAFE-NET converges faster and typically outperforms deeper networks and more complex architectures. It consistently uses fewer parameters – on average, 65% fewer than the competing feature engineering methods – while achieving comparable accuracy in less than 30% of the training epochs. Moreover, each SAFE-NET epoch is 95% faster than those of competing feature engineering approaches. These findings challenge the prevailing belief that modern PINNs effectively learn features in these scientific applications and highlight the efficiency gains possible through feature engineering.
Retrieval-augmented generation (RAG) has become a standard mechanism for grounding language models in external knowledge, yet conventional retrieval based on lexical or semantic similarity is poorly suited for complex reasoning tasks: a semantically similar problem may demand an entirely different solution strategy, while a superficially different problem may share the same underlying reasoning pattern. We propose Retrieval-Augmented Reinforcement Fine-Tuning (RA-RFT), a post-training framework that teaches language models to reason by analogy. RA-RFT uses gold-relevance distillation to train a retriever that ranks contexts by expected reasoning benefit rather than semantic overlap, and then fine-tunes the policy model via reinforcement fine-tuning methods with retrieved analogous demonstrations, so the model learns to leverage reasoning traces under verifiable outcome rewards. We further analyze the diversity of retrieved contexts and find that reasoning-aware retrieval surfaces complementary solution strategies that provide distinct reasoning scaffolds for individual problems. Across challenging mathematical reasoning benchmarks, RA-RFT consistently outperforms standard reinforcement fine-tuning methods. For example, it improves AIME 2025 average@32 accuracy by 7.1 and 2.8 points over GRPO for Qwen3-1.7B and Qwen3-4B respectively – suggesting that reasoning-aware retrieval is a complementary axis of improvement and orthogonal to advances in reward design or training curricula.
Migratory cells tend to have soft nuclei that deform and penetrate narrow spaces1,2. Extensive nuclear deformation during migration can cause nuclear-envelope rupture and DNA damage in cancer cells, which may contribute to malignant transformation during tumour progression3–6. However, the importance of DNA damage in physiological migration is less well understood. Here we demonstrate that the migration of neurons in developing cerebral and cerebellar cortices is accompanied by massive DNA double-stranded breaks (DSBs) due to mechanostress during passage through narrow interstitial spaces. In contrast to many other migratory cells, these DSBs occur without detectable nuclear envelope rupture. Confined migration increases topoisomerase-IIβ covalently bound DSBs, and these lesions are repaired through non-homologous end-joining during brain development without causing cell death. Genome sequencing revealed that DSBs tend to occur at transcriptionally inactive regions. The deletion of ligase IV at the onset of neuronal migration leads to persistent DSB accumulation in cerebellar neurons with moderate transcriptional changes in genes related to synaptic function, neuronal development and stress and immune responses. The mutant mouse develops mild motor deficits in later life, suggesting that the DNA damage generated during normal brain development poses a potential disease risk if left unrepaired. The migration of neurons in developing cerebral and cerebellar cortices is accompanied by massive DNA double-strand breaks due to mechanostress during passage through narrow interstitial spaces.
arXiv:2604.03275v2 Announce Type: replace-cross Abstract: Effective adaptation and mitigation strategies for climate change require high-resolution projections to inform strategic decision-making. Conventional global climate models, which typically operate at resolutions of 150 to 200 kilometers, lack the capacity to represent essential regional processes. IPSL-AID is a global to regional downscaling tool based on a denoising diffusion probabilistic model designed to address this limitation. Trained on ERA5 reanalysis data, it generates 0.25 degree resolution fields for temperature, wind, and precipitation using coarse inputs and their spatiotemporal context. It also models probability distributions of fine-scale features to produce plausible scenarios for uncertainty quantification. The model accurately reconstructs statistical distributions, including extreme events, power spectra, and spatial structures. This work highlights the potential of generative diffusion models for efficient climate downscaling with uncertainty
arXiv:2606.18367v1 Announce Type: new Abstract: Standard benchmarks evaluate time series foundation models (TSFMs) using aggregate metrics, but these can mask severe failures in critical operating regimes. We introduce regime-stratified evaluation and apply it to three TSFMs on two standard traffic speed benchmarks. Traffic exhibits abrupt regime switching between free-flow and congested states, producing bimodal speed distributions during transitions. When we stratify by traffic regime, both accuracy and prediction-interval coverage degrade sharply during transitions: transition-regime MAE reaches 11 mph (versus 3 mph overall), and empirical coverage of 90% prediction intervals drops as low as 55%. These failures are invisible in aggregate metrics because free-flow observations dominate the sample. A simple historical conditional baseline (sampling from per-sensor training distributions) achieves better transition coverage than any TSFM, but has far worse overall accuracy. We propose bimodal mixture augmentation (BMA), a post-hoc method that combines TSFM forecasts with historical distributional knowledge, approaching the historical baseline's transition coverage while preserving the TSFM's accuracy. Our results suggest that TSFM benchmarks should incorporate regime-aware evaluation to surface failures that aggregate metrics hide.
Pixel-space diffusion models are trained on full-bandwidth noisy images, yet the useful signal available to the denoiser is strongly frequency dependent. Under rectified-flow diffusion and natural-image power-law spectra, the per-band data-to-noise contour $k^{*}(t) = (1-t)^{-2/\alpha}$ separates a signal-bearing low-frequency region from a noise-dominated high-frequency region at each time $t$. We show that this implicit coarse-to-fine structure is not merely descriptive: it induces a capacity-allocation problem. A standard pixel-space denoiser must discover the moving bandwidth boundary internally and can spend computation on frequency-time regions where the optimal prediction collapses to deterministic baselines rather than data-distribution modeling. To make this boundary explicit, we introduce Spectral Forcing, a parameter-free, time-conditional 2D-DCT low-pass operator applied to the noisy input before the patch embedder. Its cutoff expands monotonically with the diffusion time and becomes the identity at the data endpoint. Through controlled synthetic experiments, we identify the regime in which the operator is beneficial: coarse patch tokenization and data whose high-frequency content is predominantly noise rather than essential signal. On ImageNet-256 with JiT-700M/32, Spectral Forcing consistently improves both FID and Inception Score across different training epochs, demonstrating robust gains throughout training; at finer tokenization, the spectral forcing is still competitive. We further insert the unchanged operator into SenseNova-U1, a unified text-to-image model, where it improves DPG-Bench and GenEval, showing that the input-side spectral prior transfers beyond class-conditional generation. These results suggest a route to capacity-efficient pixel-space diffusion by showing the signal and hiding the noise.
Vision Transformers (ViTs) have become a dominant architecture for visual representation learning, providing exceptionally strong and broadly reusable backbone features. However, ViTs are commonly operated on relatively small patch-token grids due to the quadratic cost of global self-attention, which creates a persistent bottleneck for dense prediction tasks such as semantic segmentation and depth estimation. This has motivated the development of task-agnostic feature upsamplers. While recent state-of-the-art methods produce visually sharp dense representations, their reliance on shallow image encoders for guided upsampling can introduce feature leakage, fragmentation, and blur. We introduce ViT-Up, an implicit feature upsampling framework that replaces external image guidance with layer-wise query construction from intermediate ViT hidden states. This enables feature prediction at arbitrary continuous image coordinates while preserving alignment with the backbone feature space. Experiments demonstrate that ViT-Up consistently outperforms state-of-the-art image-guided upsamplers across dense prediction and semantic correspondence. On DINOv3-S+, ViT-Up improves over prior methods by up to +2.07 mIoU on Cityscapes and +4.17 PCK@0.10 on SPair-71k. With the larger DINOv3-B backbone, these gains increase to +3.36 mIoU and +8.09 PCK@0.10, demonstrating that ViT-Up scales favorably with backbone capacity.
T cell receptor (TCR)-based immunotherapy holds immense potential for treating cancers and infectious diseases, where highly antigen-specific TCR recognition is crucial for adaptive immunity against tumors and pathogens. Engineering or de novo generation of the complementarity-determining region 3 (CDR3) loops of TCRs using artificial intelligence offers a powerful alternative to designing reactive TCRs rather than laborious experimental screening. However, current in silico approaches are constrained by weak conditional guidance, limited flexibility, and a lack of rigorous functional validation. To address these limitations, we introduce TCRDiff, a generative diffusion framework for designing antigen-specific TCRs conditioned on peptide-MHC (pMHC) targets and germline-encoded variable genes. By leveraging pre-trained knowledge from massive T-cell repertoires and TCR-pMHC recognition data, TCRDiff generates CDR3{beta} sequences with state-of-the-art fidelity to native binding TCRs through a denoising diffusion process. Furthermore, incorporating the interface geometry features generated TCR-pMHC complexes with superior structural plausibility. As a proof of concept, we deployed TCRDiff in a systematic pipeline to design candidate TCRs for immunotherapy. In vitro activation assays validated that TCRDiff-generated TCRs specifically recognize the MAGE-A3 epitope with minimized off-target cross-reactivity. Together, TCRDiff establishes a powerful, validated computational paradigm to accelerate the development of TCR-based immunotherapies.
arXiv:2606.18617v1 Announce Type: cross Abstract: There exist numerous tutor training platforms. However, few provide AI-driven training and evaluation for human tutors based on real-life performance. We present an AI-driven system that assesses both open responses during training and authentic real-life tutoring. Unlike platforms that only assess learning through online training or simulations, our system utilizes Generative AI (Gemini-2.5-pro) to analyze transcriptions of authentic tutoring, measuring the transfer of tutor skills to real-life application. Human tutors instructing students remotely in math (N=86) completed six scenario-based lessons, averaging a significant 7.4% learning gain. Using mixed-effects models across 405 session-to-lesson pairs, we found that training performance significantly predicted real-life transcript scores with an effect size of 0.25 SD. Model comparison (AIC/BIC) indicated averaging open response and multiple choice performance during training predicted real-life tutor performance best, although open responses were comparatively more predictive. Exploratory analysis showed that after training, tutors were significantly more likely to encounter pedagogical opportunities to apply their skills (61.1% to 68.9%) and demonstrated higher execution quality within those opportunities (65.5% to 68.1%). Interrupted time series analysis suggested that these tutor improvements were part of a gradual trend over time rather than an immediate intervention effect of training. We illustrate an AI-driven method to link tutor training with real-life assessment. In doing so, we contribute open datasets, AI prompts, and scoring rubrics to support transparency and reproducibility.
Event cameras capture dynamic scenes with exceptional temporal fidelity by representing them as a continuous stream of microsecond resolution events. Each individual event, however, only carries minimal semantic value, merely signaling a localized brightness change. To derive meaningful signals, downstream algorithms need to quickly integrate cues from a potentially massive torrent of low-information events. Current architectures, however, are easily overwhelmed, struggling to balance capturing fine-grained temporal dynamics and maintaining a manageable data throughput. This paper proposes a framework to re-tokenize event streams into a small set of highly informative neural events, each representing a local spatio-temporal context window with a discrete learnable code. Every time this code flips, a neural event is triggered, yielding a highly compressed data stream. We demonstrate that, across object detection and classification, networks trained on neural events are on par or surpass the performance of state-of-the-art approaches while reducing the event rate by a factor of 2.0.
arXiv:2605.11287v2 Announce Type: replace-cross Abstract: A persistent paradox in time-series forecasting is that structurally simple MLP and linear models often outperform high-capacity Transformers. We argue that this gap arises from a mismatch in the sequence-modeling primitive: while many time-series dynamics are governed by global temporal operators (e.g., filtering and harmonic structure), standard attention forms each output as a convex combination of inputs. This restricts its ability to represent signed and oscillatory transformations that are fundamental to temporal signal processing. We formalize this limitation as a simplex-constrained mixing bottleneck in softmax attention, which becomes especially restrictive for operator-driven time-series tasks. To address this, we propose $Temporal Operator Attention (TOA)$, a framework that augments attention with explicit, learnable sequence-space operators, enabling direct signed mixing across time while preserving input-dependent adaptivity. To make dense $N \times N$ operators practical, we introduce Stochastic Operator Regularization, a high-variance dropout mechanism that stabilizes training and prevents trivial memorization. Across forecasting, anomaly detection, and classification benchmarks, TOA consistently improves performance when integrated into standard backbones such as PatchTST and iTransformer, with particularly strong gains in reconstruction-heavy tasks. These results suggest that explicit operator learning is a key ingredient for effective time-series modeling.
arXiv:2606.17319v1 Announce Type: cross Abstract: Motivated by the optimization of bounded binary black-box functions, we study the problem of learning polynomial surrogates over the Boolean hypercube. To ensure that optimizing the surrogate yields good solutions for the underlying objective, we require uniform $L_\infty$-error guarantees rather than the usual $L_2$-type guarantees. We characterize the minimax sample complexity of uniform estimation under subgaussian noise for two classes of bounded polynomials. First, for polynomials of degree at most $d$ on $n$ variables, the sample complexity scales as $n^{d+1}$. Second, for $s$-sparse Fourier-Walsh polynomials with $s \leq n$, it scales as $ns^2$. These rates differ structurally from the noiseless setting, where uniform exact recovery scales as $n^d$ and $ns$, respectively. Our lower bounds hold even for arbitrary adaptive learners, showing that the additional factors are intrinsic to the noisy cases. Standard Fourier-analysis tools for the $L_2$-norm do not naturally extend to the $L_\infty$-setting in a way that yields uniform guarantees. Our proofs overcome this difficulty by relying on suitably chosen auxiliary norms that serve as proxies for controlling the $L_\infty$-error. Together, our results provide a tight characterization of the sample complexity of learning optimization-safe polynomial surrogates.
arXiv:2606.19057v1 Announce Type: cross Abstract: Large Language Models (LLMs) are increasingly used as judges for scalable evaluation, yet such LLM–as–a–Judge systems exhibit systematic biases that are decoupled from semantic quality, most notably verbosity bias. Meanwhile, human supervision is costly and typically selective, yielding reliable positive judgments but leaving most outputs unlabelled and potentially mixed in quality. We formulate LLM evaluation under selective human supervision as a positive–unlabelled learning problem and propose a geometric auditing framework based on Partial Optimal Transport. By aligning a small set of human–verified positives with a reliable subset of unlabelled outputs in a fixed embedding space, our method identifies human–consistent preferences and corrects biased judges without retraining. Experiments demonstrate improved alignment with human preferences, increased robustness to presentation biases, and interpretable confidence estimates, offering a scalable and statistically grounded alternative to existing LLM–as–a–judge pipelines.
Diffusion large language models (dLLMs) have emerged as a promising alternative for text generation, distinguished by their native support for parallel decoding. In practice, block inference is crucial for avoiding order misalignment in global bidirectional decoding and improving output quality. However, the widely-used fixed, predefined block (naive) schedule is agnostic to semantic difficulty, making it a suboptimal strategy for both quality and efficiency: it can force premature commitments to uncertain positions while delaying easy positions near block boundaries. In this work, we analyze the limitations of naive block scheduling and disclose the importance of dynamically adapting the schedule to semantic difficulty for reliable and efficient inference. Motivated by this, we propose Dynamic Sliding Block (DSB), a training-free block scheduling method that uses a sliding block with a dynamic size to overcome the rigidity of the naive block. To further improve efficiency, we introduce DSB Cache, a training-free KV-cache mechanism tailored to DSB. Extensive experiments across multiple models and benchmarks demonstrate that DSB, together with DSB Cache, consistently improves both generation quality and inference efficiency for dLLMs. Code is released at https://github.com/lizhuo-luo/DSB.
arXiv:2606.18256v1 Announce Type: cross Abstract: LLM-based chatbots are increasingly applied in interpersonal domains such as counseling and peer support, where establishing human-AI rapport is crucial yet remains challenging. In this work, we introduce a novel approach for conditioning LLMs with in-group personas, which (i) first identifies a user's primary concern and brief personal context (e.g., a computer science undergraduate worried about future career prospects), and (ii) generates a synthetic in-group persona that shares a similar primary concern while differing in background and narrative details, such as age or profession (e.g., a junior researcher at an AI startup). Furthermore, we conduct a human-subject study to systematically evaluate the effectiveness of in-group persona agents in enhancing human-AI rapport. We compare our approach against two baseline conditions: a conventional agent without persona conditioning and an agent exhibiting minimal self-disclosure (e.g., "I've felt that too"). Results from post-task questionnaires assessing rapport and user experience indicate that the in-group persona agent significantly improves perceived rapport and personal relevance compared to the baselines, and also yields more positive user experience-most notably higher engagement.
Reliable autonomous driving requires vectorized HD maps that are geometrically accurate, semantically rich, and scalable to long-horizon driving. However, existing public HD map datasets are limited in scale, provide sparse semantic attributes, and lack modalities such as aerial imagery that could enable new research directions. We present HRDX, a large-scale dataset for vector HD-map construction, spanning about 40 hours (1,400 km) of minimally overlapping drives, which is several times larger than prior public HD map datasets. Data is captured using six synchronized surround cameras, a 128-beam LiDAR, and centimeter-level RTK GNSS/IMU, and is further complemented by precisely aligned aerial orthoimagery. Annotations cover 10 vector map classes, complemented with over 20 semantic and topological attributes. To evaluate this richer ontology, we introduce the Composite Score (CS) to jointly assess geometric fidelity and attribute correctness. Benchmark experiments show that HRDX's scale improves online vector-map construction, and that aligned aerial imagery provides a useful structural prior: using aerial imagery at training and/or inference improves geometric map quality, while aerial-augmented teachers can transfer part of this benefit to camera-only students without increasing inference-time sensor requirements. HRDX is intended to support reproducible research on large-scale HD-map learning, multimodal BEV fusion, and training-time privileged information. HRDX dataset and benchmarks are available at https://github.com/honda-research-institute/HRDX
arXiv:2407.04884v4 Announce Type: replace Abstract: The hidden state threat model of differential privacy (DP) assumes that the adversary has access only to the final trained machine learning (ML) model, without seeing intermediate states during training. However, the current privacy analyses under this model are restricted to convex optimization problems, reducing their applicability to multi-layer neural networks, which are essential in modern deep learning applications. Notably, the most successful applications of the hidden state privacy analyses in classification tasks have only been for logistic regression models. We demonstrate that it is possible to privately train convex problems with privacy-utility trade-offs comparable to those of 2-layer ReLU networks trained with DP stochastic gradient descent (DP-SGD). This is achieved through a stochastic approximation of a dual formulation of the ReLU minimization problem, resulting in a strongly convex problem. This enables the use of existing hidden state privacy analyses and provides accurate privacy bounds also for the noisy cyclic mini-batch gradient descent (NoisyCGD) method with fixed disjoint mini-batches. Empirical results on benchmark classification tasks demonstrate that NoisyCGD can achieve privacy-utility trade-offs on par with DP-SGD applied to 2-layer ReLU networks.
arXiv:2606.08304v2 Announce Type: replace-cross Abstract: We investigate the relationship between the analytical properties of functions of bounded variation and the statistical behavior of hyperuniform point processes. We establish several characterization formulas for the jump part of the gradient of a bounded variation function, extending and unifying previous results by Beretti–Gennaioli and Dávila. In particular, we provide new expressions for the $L^2$-jump of the gradient using both difference quotients and Fourier transform methods. Furthermore, we connect these analytic structures to the theory of hyperuniform point processes. By analyzing the variance of linear statistics associated with bounded variation functions, we provide asymptotic estimates that depend on the specific classification of the hyperuniformity of the point process. The results show how the regularity and jump discontinuities of a function dictate the growth rate of fluctuations in point processes. Finally, we introduce an averaged quadratic BMO-type oscillation functional over translated and rotated cube partitions, similar to the one recently studied by Ambrosio et al., and prove, using results from point process, that it converges to an explicit dimensional constant times the $L^2-$jump, giving in particular a further new characterization of the perimeter of a set.
arXiv:2606.19683v1 Announce Type: new Abstract: This paper studies coalition formation as a decentralized dynamical process driven by unilateral exit-and-join decisions. Agents evaluate local moves using the Aumann-Dreze value, so payoffs are computed within the agent's current coalition rather than through a globally negotiated coalition structure. The resulting model links cooperative payoff allocation with noncooperative best-response behavior: a terminal partition is precisely a coalition structure with no admissible, individually profitable exit-and-join deviation. We establish equilibrium characterizations, identify conditions under which the dynamics admit scalar Lyapunov or exact-potential representations, and analyze how switching and acceptance costs shape local stability. Numerical experiments test finite-time stabilization, cost sensitivity, and a special convex-game benchmark.
arXiv:2505.20030v2 Announce Type: replace-cross Abstract: We observe a novel `multiple-descent' phenomenon during the learning process of a recurrent neural network called long-short-term memory (LSTM) networks during its training on real-world task, in which the performance goes through long cycles of up and down trends multiple times after the model is overtrained. By carrying out asymptotic stability analysis of the models, we found that the cycles in performance – indicated by loss function in test data – are closely associated with the phase transition process between order and chaos of the model, and the local optimal training step are consistently at the critical transition point between the two phases. More importantly, the most optimal point of the model usually occurs at the first transition from order to chaos, where the `width' of the `edge of chaos' is often the widest, allowing the best exploration of weight configurations for learning.
arXiv:2606.18867v1 Announce Type: new Abstract: When algorithmic predictors inform resource allocation in high-stakes domains such as healthcare, these predictors must account for strategic manipulation of input features. The typical solution is to redesign the predictor itself to explicitly account for strategic interactions. In practice, however, decision makers are often constrained to adjusting coarser levers within existing prediction pipelines. For example, healthcare organizations often select which features to exclude based on perceived manipulability, while using standard regularization procedures to shrink the coefficients of retained features. In this work, we initiate a formal study of strategic classification through feature selection and its interaction with ridge regularization. Our main finding is that excluding individual features based on their manipulability alone is generally suboptimal. We provide a fine-grained characterization of the performance of a feature subset under optimal regularization, yielding new insights for policy design. Motivated by this characterization, we develop a practical algorithm for jointly choosing the feature set and the level of ridge regularization. Through a real-world case study on a healthcare payments benchmark, we illustrate how our algorithm can guide the design of coarse policy levers in practice. Our results provide a principled, practical framework for mitigating the effects of strategic behavior in algorithmic decision-making systems.
arXiv:2602.23238v2 Announce Type: replace Abstract: Photon sources based on spontaneous parametric down-conversion (SPDC) are central to experimental quantum optics and quantum technologies. Their performance is commonly quantified by three metrics: pair-collection probability, heralding efficiency, and spectral purity. In bulk-crystal SPDC, these metrics are known to be mutually constrained, yet the physical origin of the resulting trade-offs is often obscured. We show that these trade-offs originate from the frequency-dependent population of discrete spatial modes in the SPDC emission. By performing a Laguerre-Gauss mode decomposition at each frequency component, we show how spectral-spatial non-separability impacts collection probability, heralding efficiency, and purity. We apply this framework to two widely used quasi-phase-matching configurations: collinear degenerate type-0 and type-II SPDC in periodically poled bulk crystals, and quantify how different phase-matching functions shape the spectral-spatial mode structure. In particular, for type-II SPDC we compare standard periodically poled and aperiodically poled Gaussian phase matching. We experimentally validate some of our theoretical results using spatial- and spectral-projection measurements. This spectral-spatial mode analysis provides a quantitative and predictive framework for understanding and engineering bulk-crystal photon sources, enabling systematic multi-parameter optimization beyond qualitative design guidelines.
arXiv:2510.19528v2 Announce Type: replace-cross Abstract: We investigate the fundamental problem of leveraging offline data to accelerate online reinforcement learning - a direction with strong potential but limited theoretical grounding. Our study centers on how to learn and apply value envelopes within this context. To this end, we introduce a principled two-stage framework: the first stage uses offline data to derive upper and lower bounds on value functions, while the second incorporates these learned bounds into online algorithms. Our method extends prior work by decoupling the upper and lower bounds, enabling more flexible and tighter approximations. In contrast to approaches that rely on fixed shaping functions, our envelopes are data-driven and explicitly modeled as random variables, with a filtration argument ensuring independence across phases. The analysis establishes high-probability regret bounds determined by two interpretable quantities, thereby providing a formal bridge between offline pre-training and online fine-tuning. Empirical results on tabular MDPs demonstrate substantial regret reductions compared with both UCBVI and prior methods while remaining competitive with related approaches.
Capsid engineering has greatly improved the performance of recombinant AAV vectors used for gene therapy. One commonly used strategy is the insertion of a short, 7-mer, peptide into surface-exposed loops to modify receptor interactions and enhance cell entry. While effective in receptor retargeting and improved transduction, these insertions might destabilize the capsid protein, hinder assembly, and thus limit production. While previous attempts have used deep mutational scanning and AI to predict which insertions are viable, there is lack in understanding the structural consequences of these peptide insertions at the amino-acid level. Here we combined experiments, deep sequencing and large protein language models to gain insight on the impact of 7-mer insertions on the VR-VIII region. We first characterize the biochemical properties of viable insertions, thus identifying which residues are well tolerated, and which should instead be avoided. We then focus on the nearby context of those insertions, by studying the effect of the linkers, either for highly diverse libraries or for individual variants known for their efficiency. Next, we study the broader context, by extending our analysis to the whole capsid sequence, and identifying regions that can tolerate insertions without long-ranged structural deformations that could affect capsid functionality. We conclude with a cross-serotype comparison and a viability analysis of tens of previously engineered variants. Our work showcases how AI can uncover structure-function rules governing the success of engineered AAV capsids.