scTOP: physics-inspired order parameters for cellular identification and visualization.

Advances in single-cell RNA sequencing provide an unprecedented window into cellular identity. The abundance of data requires new theoretical and computational frameworks to analyze the dynamics of differentiation and integrate knowledge from cell atlases. We present 'single-cell Type Order Parameters' (scTOP): a statistical, physics-inspired approach for quantifying cell identity given a reference basis of cell types. scTOP can accurately classify cells, visualize developmental trajectories and assess the fidelity of engineered cells. Importantly, scTOP does this without feature selection, statistical fitting or dimensional reduction (e.g. uniform manifold approximation and projection, principle components analysis, etc.). We illustrate the power of scTOP using human and mouse datasets. By reanalyzing mouse lung data, we characterize a transient hybrid alveolar type 1/alveolar type 2 cell population. Visualizations of lineage tracing hematopoiesis data using scTOP confirm that a single clone can give rise to multiple mature cell types. We assess the transcriptional similarity between endogenous and donor-derived cells in the context of murine pulmonary cell transplantation. Our results suggest that physics-inspired order parameters can be an important tool for understanding differentiation and characterizing engineered cells. scTOP is available as an easy-to-use Python package.

Defining developmental trajectories of prosensory cells in human inner ear organoids at single-cell resolution.

The inner ear sensory epithelia contain mechanosensitive hair cells and supporting cells. Both cell types arise from SOX2-expressing prosensory cells, but the mechanisms underlying the diversification of these cell lineages remain unclear. To determine the transcriptional trajectory of prosensory cells, we established a SOX2-2A-ntdTomato human embryonic stem cell line using CRISPR/Cas9, and performed single-cell RNA-sequencing analyses with SOX2-positive cells isolated from inner ear organoids at various time points between differentiation days 20 and 60. Our pseudotime analysis suggests that vestibular type II hair cells arise primarily from supporting cells, rather than bi-fated prosensory cells in organoids. Moreover, ion channel- and ion-transporter-related gene sets were enriched in supporting cells versus prosensory cells, whereas Wnt signaling-related gene sets were enriched in hair cells versus supporting cells. These findings provide valuable insights into how prosensory cells give rise to hair cells and supporting cells during human inner ear development, and may provide a clue to promote hair cell regeneration from resident supporting cells in individuals with hearing loss or balance disorders.

Longitudinal mapping of the development of cortical thickness and surface area in rhesus macaques during the first three years.

Studying dynamic spatiotemporal patterns of early brain development in macaque monkeys is critical for understanding the cortical organization and evolution in humans, given the phylogenetic closeness between humans and macaques. However, due to huge challenges in the analysis of early brain Magnetic Resonance Imaging (MRI) data typically with extremely low contrast and dynamic imaging appearances, our knowledge of the early macaque cortical development remains scarce. To fill this critical gap, this paper characterizes the early developmental patterns of cortical thickness and surface area in rhesus macaques by leveraging advanced computing tools tailored for early developing brains based on a densely sampled longitudinal dataset with 140 rhesus macaque MRI scans seamlessly covering from birth to 36 mo of age. The average cortical thickness exhibits an inverted U-shaped trajectory with peak thickness at around 4.3 mo of age, which is remarkably in line with the age of peak thickness at 14 mo in humans, considering the around 3:1 age ratio of human to macaque. The total cortical surface area in macaques increases monotonically but with relatively lower expansions than in humans. The spatial distributions of thicker and thinner regions are quite consistent during development, with gyri having a thicker cortex than sulci. By 4 mo of age, over 81% of cortical vertices have reached their peaks in thickness, except for the insula and medial temporal cortices, while most cortical vertices keep expanding in surface area, except for the occipital cortex. These findings provide important insights into early brain development and evolution in primates.

Single-nuclei transcriptome analysis of the shoot apex vascular system differentiation in Populus.

Differentiation of stem cells in the plant apex gives rise to aerial tissues and organs. Presently, we lack a lineage map of the shoot apex cells in woody perennials - a crucial gap considering their role in determining primary and secondary growth. Here, we used single-nuclei RNA-sequencing to determine cell type-specific transcriptomes of the Populus vegetative shoot apex. We identified highly heterogeneous cell populations clustered into seven broad groups represented by 18 transcriptionally distinct cell clusters. Next, we established the developmental trajectories of the epidermis, leaf mesophyll and vascular tissue. Motivated by the high similarities between Populus and Arabidopsis cell population in the vegetative apex, we applied a pipeline for interspecific single-cell gene expression data integration. We contrasted the developmental trajectories of primary phloem and xylem formation in both species, establishing the first comparison of vascular development between a model annual herbaceous and a woody perennial plant species. Our results offer a valuable resource for investigating the principles underlying cell division and differentiation conserved between herbaceous and perennial species while also allowing us to examine species-specific differences at single-cell resolution.

CProtMEDIAS: clustering of amino acid sequences encoded by gene families by MErging and DIgitizing Aligned Sequences.

Protein phylogenetic analysis focuses on the evolutionary relationships among related protein sequences and can help researchers infer protein functions and developmental trajectories. With the advent of the big data era, the existing protein phylogenetic methods, including distance matrix and character-based methods, are facing challenges in both running time and application scope. Here, we developed an R package that we call CProtMEDIAS that is useful for protein phylogenetic analysis. In contrast to existing phylogenetic analysis methods, CProtMEDIAS utilizes dimensionality reduction algorithms to digitize multiple sequence alignments and quickly conduct phylogenetic analysis with a large number of amino acid sequences from similarly distant protein families and species. We used CProtMEDIAS to perform a dimensionality reduction, clustering, pseudotime, specific residue and evolutionary trajectory analysis of the plant homeobox superfamily. We found that CProtMEDIAS delivers consistent clustering, fast running and elegant presentation and thus provides powerful new tools and methods for protein clustering and evolutionary analysis.

Unraveling GRIA1 neurodevelopmental disorders: Lessons learned from the p.(Ala636Thr) variant.

Ionotropic glutamate receptors (iGluRs), specifically α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptors (AMPARs), play a crucial role in orchestrating excitatory neurotransmission in the brain. AMPARs are intricate assemblies of subunits encoded by four paralogous genes: GRIA1-4. Functional studies have established that rare GRIA variants can alter AMPAR currents leading to a loss- or gain-of-function. Patients affected by rare heterozygous GRIA variants tend to have family specific variants and only few recurrent variants have been reported. We deep-phenotyped a cohort comprising eight unrelated children and adults, harboring a recurrent and well-established disease-causing GRIA1 variant (NM_001114183.1: c.1906G>A, p.(Ala636Thr)). Recurrent symptoms included motor and/or language delay, mild-severe intellectual disability, behavioral and psychiatric comorbidities, hypotonia and epilepsy. We also report challenges in social skills, autonomy, living and work situation, and occupational levels. Furthermore, we compared their clinical manifestations in relation to those documented in patients presenting with rare heterozygous variants at analogous positions within paralogous genes. This study provides unprecedented details on the neurodevelopmental outcomes, cognitive abilities, seizure profiles, and behavioral abnormalities associated with p.(Ala636Thr) refining and broadening the clinical phenotype.

Developmental trajectory of depressive symptoms among left-behind adolescents: The effects of parent-adolescent separation and parent-adolescent cohesion.

INTRODUCTION: Left-behind adolescents are vulnerable to depressive symptoms under the context of parent-adolescent separation. However, limited knowledge is available regarding left-behind adolescents' depression trajectory and the protective resources against it. The aim of this longitudinal study was to investigate the depression trajectory and its association with parent-adolescent separation (left-behind status, age of separation and duration of separation) and parent-adolescent cohesion (father-adolescent cohesion, mother-adolescent cohesion) among left-behind adolescents. METHODS: The participants were 1,107 left-behind adolescents (Mage = 13.23 ± 0.86 years at T1; 45.17% girls; 38.48% both-parent migrant adolescents, 61.52% father-only migrant adolescents) from two rural areas of Shandong Province in China, who were participated in assessment at three time points with 6 months apart from November 2014 to November 2015. RESULTS: The results indicated that the trajectory of left-behind adolescents' depression symptoms showed a decreasing trend. Both-parent migrant adolescents reported a higher initial level of depressive symptoms than father-only migrant adolescents. Duration of separation positively predicted the initial level of left-behind adolescents' depressive symptoms. Mother-adolescent cohesion negatively predicted the initial level and positively predicted the change rate of left-behind adolescents' depressive symptoms. Moreover, mother-adolescent and father-adolescent cohesion buffered the negative effect of parent-adolescent separation on the initial level of left-behind adolescents' depressive symptoms. CONCLUSIONS: These findings highlight the protective role of parent-adolescent cohesion for left-behind adolescents, having important implications for interventions targeted toward mitigating the detrimental influence of parent-adolescent separation on left-behind adolescents' depression trajectory.

The developmental trajectory of prosocial behavior in economically disadvantaged children: General tendencies and heterogeneity.

INTRODUCTION: This study explored the general tendencies and heterogeneous developmental trajectory of prosocial behavior and predictors. METHOD: The present study conducted latent growth model and growth mixture model analyses in a sample of 814 students (Mage = 13.79 years old at baseline; 57% girls) from economically disadvantaged families, classified as being below the local income threshold in China, with four follow-up surveys administered during the following 2 years. RESULTS: The general tendency in the developmental trajectory of prosocial behavior showed a linear decrease. A gender difference in initial levels was observed, with girls showing a higher initial level of prosocial behavior than boys. Family functioning, subjective support, and support utilization significantly affected the intercept, but objective support significantly negatively affected the slope. Heterogeneity in the development of prosocial behavior was best classified with a 3-class solution, including C1 (Rapid-decrease, 10.6%), C2 (Medium-stable, 42.5%), and C3 (High-increase, 46.9%). The patterns of prosocial behavior development in economically disadvantaged children with higher family functioning were more likely to be in the High-increase Class than in the Rapid-decrease Class. CONCLUSION: The present study revealed an average decline in the trajectories of prosocial behavior development in economically disadvantaged children. However, it also captured heterogeneous developmental trajectories. Furthermore, the study revealed that family functioning, subjective support, and support utilization all served as protective factors for prosocial behavior among economically disadvantaged children.

Trajectory patterns and influencing factors of supportive care needs in stroke patients: A longitudinal study.

AIM: To investigate the trajectory patterns and influencing factors of supportive care needs in stroke patients. DESIGN: A longitudinal study. METHODS: In total, 207 stroke patients who received treatment at the Department of Neurology in a hospital in Xuzhou between July 2022 and July 2023 were recruited using convenience sampling. Questionnaires including supportive care needs, hospital anxiety and depression scale, and the Barthel index were investigated at baseline and at 1, 3, and 6 months. A latent class growth model was applied to identify the supportive care needs trajectories. Multiple logistic regression was used to determine the predictors for membership. This study adheres to STROBE reporting guidelines. RESULTS: Three patterns of supportive care needs trajectories were identified: A high needs slow decline group (20.8%), a medium needs stable group (56.5%) and a medium needs rapid decline group (22.7%). Based on further analysis, the findings indicated that age, education level, monthly income, comorbidity, activities of daily living, anxiety and depression were associated with the trajectory categories of supportive care needs with stroke patients. CONCLUSION: This study demonstrates heterogeneity in changes in supportive care needs among stroke patients. Healthcare providers need to consider these different categories of needs and develop individualized care measures based on the characteristics of different patients. IMPACT: Healthcare providers should be aware of the fluctuations in care needs of stroke patients at various stages. Additionally, the study aimed to identify patients' specific needs based on their circumstances, monitor the rehabilitation process and establish a more personalized and optimized care plan through multidisciplinary collaboration. The ultimate goal was to alleviate symptomatic distress and address the long-term care needs of patients. PATIENT OR PUBLIC CONTRIBUTION: No patient or public contribution.

Online Self-Disclosure and Self-Concept Clarity Among Chinese Middle School Students: A Longitudinal Study.

The relationship between online self-disclosure and self-concept clarity has been previously examined through cross-sectional studies. This study examined causal connections between online self-disclosure and self-concept clarity among Chinese middle school students using longitudinal data collected over 18 months. Participants were 535 seventh-grade students aged 12-14 years (Mage = 12.93, SD = 0.54, 43.18% girls), assessed four times, six months apart. In a random intercept cross-lagged panel model, self-concept clarity significantly predicted online self-disclosure. Latent growth mixture modeling identified two distinct growth trajectories for both online self-disclosure (Rapid change, 7%; Slow change, 93%) and self-concept clarity (Rapid change, 8%; No change, 92%). Multiple logistic regression analysis suggested that changes in self-concept clarity influenced the developmental trajectory profile of online self-disclosure. Although male and female students differed in online self-disclosure and self-concept clarity, gender differences in the developmental trajectory profiles of online self-disclosure and self-concept clarity were not significant. Supporting adolescents in developing a clear self-concept to mitigate risks associated with excessive online self-disclosure is important.

eSPRESSO: topological clustering of single-cell transcriptomics data to reveal informative genes for spatio-temporal architectures of cells.

BACKGROUND: Bioinformatics capability to analyze spatio-temporal dynamics of gene expression is essential in understanding animal development. Animal cells are spatially organized as functional tissues where cellular gene expression data contain information that governs morphogenesis during the developmental process. Although several computational tissue reconstruction methods using transcriptomics data have been proposed, those methods have been ineffective in arranging cells in their correct positions in tissues or organs unless spatial information is explicitly provided. RESULTS: This study demonstrates stochastic self-organizing map clustering with Markov chain Monte Carlo calculations for optimizing informative genes effectively reconstruct any spatio-temporal topology of cells from their transcriptome profiles with only a coarse topological guideline. The method, eSPRESSO (enhanced SPatial REconstruction by Stochastic Self-Organizing Map), provides a powerful in silico spatio-temporal tissue reconstruction capability, as confirmed by using human embryonic heart and mouse embryo, brain, embryonic heart, and liver lobule with generally high reproducibility (average max. accuracy = 92.0%), while revealing topologically informative genes, or spatial discriminator genes. Furthermore, eSPRESSO was used for temporal analysis of human pancreatic organoids to infer rational developmental trajectories with several candidate 'temporal' discriminator genes responsible for various cell type differentiations. CONCLUSIONS: eSPRESSO provides a novel strategy for analyzing mechanisms underlying the spatio-temporal formation of cellular organizations.

Cellular diversity and lineage trajectory: insights from mouse single cell transcriptomes.

Single cell RNA-sequencing (scRNA-seq) technology has matured to the point that it is possible to generate large single cell atlases of developing mouse embryos. These atlases allow the dissection of developmental cell lineages and molecular changes during embryogenesis. When coupled with single cell technologies for profiling the chromatin landscape, epigenome, proteome and metabolome, and spatial tissue organisation, these scRNA-seq approaches can now collect a large volume of multi-omic data about mouse embryogenesis. In addition, advances in computational techniques have enabled the inference of developmental lineages of differentiating cells, even without explicitly introduced genetic markers. This Spotlight discusses recent advent of single cell experimental and computational methods, and key insights from applying these methods to the study of mouse embryonic development. We highlight challenges in analysing and interpreting these data to complement and expand our knowledge from traditional developmental biology studies in relation to cell identity, diversity and lineage differentiation.

On tumoural growth and treatment under cellular dedifferentiation.

Differentiated cancer cells may regain stem cell characteristics; however, the effects of such a cellular dedifferentiation on tumoural growth and treatment are currently understudied. Thus, we here extend a mathematical model of cancer stem cell (CSC) driven tumour growth to also include dedifferentiation. We show that dedifferentiation increases the likelihood of tumorigenesis and the speed of tumoural growth, both modulated by the proliferative potential of the non-stem cancer cells (NSCCs). We demonstrate that dedifferentiation also may lead to treatment evasion, especially when a treatment solely targets CSCs. Conversely, targeting both CSCs and NSCCs in parallel is shown to be more robust to dedifferentiation. Despite dedifferentiation, perturbing CSC-related parameters continues to exert the largest relative effect on tumoural growth; however, we show the existence of synergies between specific CSC- and NSCC-directed treatments which cause superadditive reductions of tumoural growth. Overall, our study demonstrates various effects of dedifferentiation on growth and treatment of tumoural lesions, and we anticipate our results to be helpful in guiding future molecular and clinical research on limiting tumoural growth in vivo.

Developmental emergence of holistic processing in word recognition.

Holistic processing (HP) of faces refers to the obligatory, simultaneous processing of the parts and their relations, and it emerges over the course of development. HP is manifest in a decrement in the perception of inverted versus upright faces and a reduction in face processing ability when the relations between parts are perturbed. Here, adopting the HP framework for faces, we examined the developmental emergence of HP in another domain for which human adults have expertise, namely, visual word processing. Children, adolescents, and adults performed a lexical decision task and we used two established signatures of HP for faces: the advantage in perception of upright over inverted words and nonwords and the reduced sensitivity to increasing parts (word length). Relative to the other groups, children showed less of an advantage for upright versus inverted trials and lexical decision was more affected by increasing word length. Performance on these HP indices was strongly associated with age and with reading proficiency. Also, the emergence of HP for word perception was not simply a result of improved visual perception over the course of development as no group differences were observed on an object decision task. These results reveal the developmental emergence of HP for orthographic input, and reflect a further instance of experience-dependent tuning of visual perception. These results also add to existing findings on the commonalities of mechanisms of word and face recognition. RESEARCH HIGHLIGHTS: Children showed less of an advantage for upright versus inverted trials compared to adolescents and adults. Relative to the other groups, lexical decision in children was more affected by increasing word length. Performance on holistic processing (HP) indices was strongly associated with age and with reading proficiency. HP emergence for word perception was not due to improved visual perception over development as there were no group differences on an object decision task.

Child and adolescent development of the brain oscillatory activity during a working memory task.

The developmental trajectories of brain oscillations during the encoding and maintenance phases of a Working Memory (WM) task were calculated. The Delayed-Match-to-Sample Test (DMTS) was applied to 239 subjects of 6-29 years, while EEG was recorded. The Event-Related Spectral Perturbation (ERSP) was obtained in the range between 1 and 25 Hz during the encoding and maintenance phases. Behavioral parameters of reaction times (RTs) and response accuracy were simultaneously recorded. The results indicate a myriad of transient and sustained bursts of oscillatory activity from low frequencies (1 Hz) to the beta range (up to 19 Hz). Beta and Low-frequency ERSP increases were prominent in the encoding phase in all age groups, while low-frequency ERSP indexed the maintenance phase only in children and adolescents, but not in late adolescents and young adults, suggesting an age-dependent neural mechanism of stimulus trace maintenance. While the latter group showed Beta and Alpha indices of anticipatory attention for the retrieval phase. Mediation analysis showed an important role of early Delta-Theta and late Alpha oscillations for mediation between age and behavioral responses performance. In conclusion, the results show a complex pattern of oscillatory bursts during the encoding and maintenance phases with a consistent pattern of developmental changes.

Ontogenesis of sociability of wild immature capuchin monkeys (Sapajus libidinosus): Sex more than personality explains interindividual variation.

Sociability is a fundamental trait that social animals need to survive and reproduce in societies. Sociability predicts how an individual can consistently interact with its conspecifics across time and situations. By studying capuchin monkeys (Sapajus libidinosus), a neotropical primate with complex social behavior and high cognitive capacity, our research aims to analyze the development of the social axis of personality of immature individuals, from birth to the third year of life. We studied wild monkeys belonging to a group with infants, juveniles, and adults of both sexes that inhabits northeastern Brazil. We analyzed the behavior of 12 immature capuchins (6 males and 6 females) in 94 h of videos recorded weekly from birth until 36 months, through daily focal sampling. We verified whether there was intraindividual consistency throughout development by fitting regression models for the effect of age on initiating affiliative social behaviors, controlling for monkey identity and sex. Our results indicate that the individuals of this study exhibit high variation in the initiation of behaviors at the beginning of infancy; there was low repeatability and high intra-individual variation during the first 3 years of life of these individuals, indicating that the social personality is not consolidated in this period. Immature females were more sociable than immature males. Therefore, differences in sociability in early life of bearded capuchin monkeys are best explained by sex rather than personality. We suggest that the high initial behavioral variation in the social axis of personality allows for plasticity influenced by the environment throughout development. The high sociability of females in infancy may be related to female philopatry and their high sociability in adulthood.

Chromatin accessibility illuminates single-cell regulatory dynamics of rice root tips.

BACKGROUND: Root development and function have central roles in plant adaptation to the environment. The modification of root traits has additionally been a major driver of crop performance since the green revolution; however, the molecular underpinnings and the regulatory programmes defining root development and response to environmental stress remain largely unknown. Single-cell reconstruction of gene regulatory programmes provides an important tool to understand the cellular phenotypic variation in complex tissues and their response to endogenous and environmental stimuli. While single-cell transcriptomes of several plant organs have been elucidated, the underlying chromatin landscapes associated with cell type-specific gene expression remain largely unexplored. RESULTS: To comprehensively delineate chromatin accessibility during root development of an important crop, we applied single-cell ATAC-seq (scATAC-seq) to 46,758 cells from rice root tips under normal and heat stress conditions. Our data revealed cell type-specific accessibility variance across most of the major cell types and allowed us to identify sets of transcription factors which associate with accessible chromatin regions (ACRs). Using root hair differentiation as a model, we demonstrate that chromatin and gene expression dynamics during cell type differentiation correlate in pseudotime analyses. In addition to developmental trajectories, we describe chromatin responses to heat and identify cell type-specific accessibility changes to this key environmental stimulus. CONCLUSIONS: We report chromatin landscapes during rice root development at single-cell resolution. Our work provides a framework for the integrative analysis of regulatory dynamics in this important crop organ at single-cell resolution.

Comprehension of indirect answers: Developmental trajectory for preschool- and early elementary school-aged children with typical development.

Indirect answers are a common type of non-literal language that do not provide an explicit "yes" or "no" to a question (e.g., "I have to work late" indirectly answered "Are you going to the party?" with a negative response). In the current study, we examined the developmental trajectory of comprehension of indirect answers among 5- to 10-year-old children with typical development. Forty-eight children, 23 boys and 25 girls, between the ages of 5 years; 0 months and 10 years; 11 months (M = 8;2, SD = 19.77 months) completed an experimental task to judge whether a verbally presented indirect answer meant yes or no (Comprehension Task) and then explain their choice (Explanation Task). Responses were scored for accuracy and coded for error analysis. On the Comprehension Task, the 5- to 8-year-olds performed with approximately 85% accuracy, while the 9- and 10-year-olds achieved 95% accuracy. On the Explanation Task, the cross-sectional trajectory revealed three stages: the 5- and 6-year-olds adequately explained indirect answers 32% of the time, the 7- and 8-year-olds performed significantly higher at 55%, and the 9- and 10-year-olds made significant gains than the younger children at 66%. Error analysis revealed that when children fail to interpret speaker intentions appropriately, they repeat the speaker's utterance or provide an insufficient explanation 80% of the time. Other responses, such as those irrelevant to the context, indicating "I don't know" or no response, or that were made-up interpretations each accounted for 2%-10% of total inadequate explanations. Study findings indicate discrepancies between task performances and offer two separate sets of baseline data for future comparisons that investigate comprehension or explanation of indirect answers by children with different cultural and linguistic backgrounds and by those with varying cognitive and language profiles.

Mapping developmental paths of monkey primordial germ-like cells differentiation from pluripotent stem cells by single cell ribonucleic acid sequencing analysis†.

The induction of primordial germ-like cells (PGCLCs) from pluripotent stem cells (PSCs) provides a powerful system to study the cellular and molecular mechanisms underlying germline specification, which are difficult to study in vivo. The studies reveal the existence of a species-specific mechanism underlying PGCLCs between humans and mice, highlighting the necessity to study regulatory networks in more species, especially in primates. Harnessing the power of single-cell RNA sequencing (scRNA-seq) analysis, the detailed trajectory of human PGCLCs specification in vitro has been achieved. However, the study of nonhuman primates is still needed. Here, we applied an embryoid body (EB) differentiation system to induce PGCLCs specification from cynomolgus monkey male and female PSCs, and then performed high throughput scRNA-seq analysis of approximately 40 000 PSCs and cells within EBs. We found that EBs provided a niche for PGCLCs differentiation by secreting growth factors critical for PGCLC specification, such as bone morphogenetic protein 2 (BMP2), BMP4, and Wnt Family Member 3. Moreover, the developmental trajectory of PGCLCs was reconstituted, and gene expression dynamics were revealed. Our study outlines the roadmap of PGCLC specification from PSCs and provides insights that will improve the differentiation efficiency of PGCLCs from PSCs.

Influences on the trajectory and subsequent outcomes in CDKL5 deficiency disorder.

OBJECTIVE: The study investigated the effect of seizure and medication burden at initial contact with the International CDKL5 Disorder Database on subsequent development and clinical severity and compared quality of life among those whose development progressed, remained stable, or regressed between baseline and follow-up. METHODS: The effects of seizure and medication burden at baseline (high or low) on the CDKL5 Disorder Severity Scores and CDKL5 Developmental Score (CDS) at follow-up were assessed using linear and negative binomial regressions, respectively, with adjustment for age at baseline, gender, and follow-up duration with and without genotype. Seizure and medication burden were defined by average daily seizure count (high, ≥5/day; low, <5/day) and number of antiseizure medications (high, ≥3/day; low, <3/day), respectively. The effects of change in CDS over time (improved, stable, or deteriorated) on Quality of Life Inventory-Disability (QI-Disability) total and domain scores at follow-up were assessed in those aged at least 3 years at follow-up using linear regression models with adjustment for baseline CDS, gender, and follow-up duration. RESULTS: The expected follow-up CDS was lower for individuals with high compared to low seizure burden at baseline (ß = -.49, 95% confidence interval [CI] = -.84 to -.13). The average total QI-Disability score was 5.6 (95% CI = -.2 to 11.5) points higher among those with improved compared with stable or deteriorating CDS and 8.5 (95% CI = 3.1-13.8) points lower for those with deteriorating compared to stable or improved CDS. SIGNIFICANCE: Our finding that later development showed slight improvement in those with better earlier seizure control even after adjustment for genotype suggests that the trajectory for an individual child is not necessarily predetermined and could possibly be influenced by optimal seizure management. This has implications for children's quality of life.