## 5 Discussion
### Limitations  
Our study does have some important limitations. The preterm and term CHD cohorts that we studied were heterogenous related to heart lesion subtypes. Also, neurodevelopmental outcomes were only available for a small subset of our primary cohort. During this study, only a limited number of socio-demographic factors were prospectively collected, and therefore analysis using these factors was unable to be carried out and future work correlating BDS score with pediatric/adolescent neurocognitive outcomes in relation to socio-demographic factors is needed. Our multi-site approach with different scanner platforms and vendors can lead to interscanner variance, but our secondary analyses did not find a correlation between scanner vendor or field strength and brain dysplasia score. Future work employing retrospective machine learning techniques like the empirical Bayes technique (Combat) can be used to reduce interscanner variance. Importantly, we were not able to measure extra-axial CSF fluid in the ECM imaging of the mouse mutants, which is an important component of the BDS score. Complete removal of CSF and infiltration of paraffin could have altered CSF volumes or the ventricle shape. With this in mind, we did not feel comfortable labeling these brain regions as containing CSF and instead labeled them as extra-axial space. As such, some of the preclinical brain regions were omitted from the analysis, resulting in missing data points. The following supporting information can be downloaded at [[https://www.mdpi.com/article/10.3390/jcm13195772/s1](https://www.mdpi.com/article/10.3390/jcm13195772/s1)]([https://www.mdpi.com/article/10.3390/jcm13195772/s1](https://www.mdpi.com/article/10.3390/jcm13195772/s1)): Table S1: Incidence of Brain Injury in Human Infant Term and Preterm CHD, Table S2: Comparison of Cortical Maturation Score (TMS) between CHD and Controls Infants (Preterm and Term), Table S3: Inter-rater Reliability (Kappa) for Brain Dysplasia Score Between Two Pediatric Neuroradiologists, Table S4A: Association Between Innate Factors, Heart Lesion Subtypes and Brain Dysplasia Score in Term CHD, Table S4B: Association Between Preoperative Risk Factors and Brain Dysplasia Score, Table S4C: Human Infant Association Between Intraoperative Factors and Brain Dysplasia Score, Table S4D: Human Association Between Post-Operative Clinical Risk Factors and Brain Dysplasia Score, Table S5: Association Between Field Strength and Brain Dysplasia Score, Table S6: Brain Dysplasia Score within _Ohiia_ Mouse Mutant Cohort: Comparison between Cardiac Lesion Subgroups, Table S7: _Ohiia_ Mouse Mutant Cohort Regional Cerebral Volumes Compared to Wild Type (WT), Table S8: Non-normalized Regional Cerebral Volumes of _Ohiia_ CHD cohorts: Comparison of Cardiac Lesion Subgroups, Table S9: Normalized Regional Cerebral Volumes of _Ohiia_ CHD cohorts: Comparison of Cardiac Lesion Subgroups, Table S10: Overview of _Ohiia_ Mutant Model Genotype Groups Studies: Congenital heart disease lesions and Holoprosencephaly Incidence, Table S11: Genotype/Brain Dysplasia Score, Table S12: Genotype/Regional Brain Volumes, Table S13: Genotype/Regional Brain Volume, Figure S1: Components of Human Brain Dysplasia Scoring Criteria, Figure S2: Components of Composite BrainInjury Score, Figure S3: Flow Diagram for NeBSS, a Semi-automated Neonatal Segmentation Pipeline, Figure S4: Flow Diagram of Sample Preparation, ECM Analysis, and Volumetric Analysis, Figure S5: Necropsy Among Mutant Mice Showed Variety and Differing Degree of Severity of Malformations, Figure S6: Individual Structures Showed a Range of Abnormality in Screen Mice, Figure S7: Varying Degree of Cerebral Abnormalities Present in Screen Mice, Figure S8: Crispr/Cas Figure Mutant Animals Showed Similar Phenotype to Screen Mice, Figure S9: Holoprosencephaly and Hippocampal Malformations Seen in Human and Mouse. Conceptualization: W.T.R., J.K.V.-S., G.G., V.K.L. and V.S. ; Data Curation: W.T.R., J.K.V.-S., G.G., Y.W., X.L., R.S., B.G., N.N.T., S.S., J.W., V.S., A.P., R.C. and C.W.L.