Search bioRxiv⌕ Search

bioRxiv · 10.1101/2023.05.17.541071

Model Parameter identification using 2D vs 3D experimental data: a comparative analysis

Abstract

Computational models are becoming an increasingly valuable tool in biomedical research. They enable the quantification of variables difficult to measure experimentally, an increase in the spatio-temporal resolution of the experiments and the testing of hypotheses. Parameter estimation from in-vitro data, remains a challenge, due to the limited availability of experimental datasets acquired in directly comparable conditions. While the use of computational models to supplement laboratory results contributes to this issue, a more extensive analysis of the effect of incomplete or inaccurate data on the parameter optimization process and its results is warranted. To this end, we compared the results obtained from the same in-silico model of ovarian cancer cell growth and metastasis, calibrated with datasets acquired from two different experimental settings: a traditional 2D monolayer, and 3D cell culture models. The differential behaviour of these models will inform the role and importance of experimental data in the calibration of computational models calibration. This work will also provide a set of general guidelines for the comparative testing and selection of experimental models and protocols to be used for parameter optimization in computational models Author summaryParameter identification is a key step in the development of a computational model, that is used to establish a connection between the simulated and experimental results and verify the accuracy of the in-silico framework. The selection of the in-vitro data to be used in this phase is fundamental, but little attention has been paid to the role of the experimental model in this process. To bridge this gap we present a comparative analysis of the same computational model calibrated using experimental data acquired from cells cultured (i) in 2D monolayers, (ii) in 3D culture models and (iii) a combination of the two. Data acquired in different experimental settings induce changes in the optimal parameter sets and the corresponding computational models behaviour. This translates in a varying degree of accuracy during the validation procedure, when the simulated data are compared to experimental measurements not used during the calibration step. Overall, our work provides a workflow and a set of guidelines to select the most appropriate experimental setting for the calibration and validation of computational models.

Source connections

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Cortesi, M., Liu, D., Yee, C., Marsh, D. J., Ford, C. E.. 2023-05-18. Model Parameter identification using 2D vs 3D experimental data: a comparative analysis. https://doi.org/10.1101/2023.05.17.541071

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

Comparative study of chlorophyll measurement in Physcomitrium patens moss using a conventional microscope adapted for combined 2D+1D imaging and spectral analysis

Imaging spectroscopy often requires expensive and complex equipment. Here we show a simple procedure for attaching a standard miniature fiber spectrometer to a conventional microscope, allowing easy integration of 2D imaging with 1D high-resolution spectral measurements. This combination provides much of the benefit of a full imaging spectrometer without the large equipment investment, and we provide instructions for modifying microscopes to this setup and the present measurements of living cells that demonstrate their performance. Using this setup, we compare the quantitative measurement of chlorophyll concentration in Physcomitrium patens moss using color imaging and spectral sampling.

bioengineering↗

De novo designed single-domain antibodies protect against lethal cobra venom neurotoxicity in vivo

Generative protein design can now rapidly produce de novo binders with high affinity and functional activity against a wide range of targets, including lethal snake venom toxins. However, so far most reported successes rely on new-to-nature scaffolds with limited therapeutic precedent. Single-domain antibodies (VHHs) offer a clinically validated alternative scaffold that can bind and neutralize long-chain -neurotoxins, which are some of the most lethal components in snake venoms. Here we compare three recently established de novo design models with VHH-design capabilities (Germinal, RFantibody, and BoltzGen) for their ability to generate VHHs against the neurotoxin -cobratoxin from the monocled cobra (Naja kaouthia). Using standardized model inputs and evaluation criteria based on AlphaFold3 interface confidence (ipTM) and RMSD self-consistency, we find that Germinal was the only method to generate designs passing stringent in silico criteria for experimental testing. We therefore performed a larger Germinal design campaign employing three different VHH frameworks and experimentally validated 46 designs in vitro. Of these, 42 expressed as soluble proteins and we identified four binding hits derived from two of the three tested frameworks. Of the four binders, two lead candidates were further characterized and demonstrated high affinity (KDs of 4.1 nM and 10.8 nM), monomeric behavior and low polyreactivity, indicating favorable biophysical and developability properties, as well as functional toxin neutralization in vitro. To assess their therapeutic potential we investigated their ability to protect against -cobratoxin toxicity in vivo. Both candidates fully protected mice after -cobratoxin challenge, with 100% survival compared to a lethal control. One candidate also retained notable neutralization capacity against whole venom of Naja kaouthia with a survival of 56%, while the other protected 22% when tested in a rescue setting. Together, we demonstrate that de novo VHH design can generate high affinity single-domain antibodies with in vivo protection against lethal cobra venom neurotoxicity, and provide practical insights into method- and framework-dependent performance.

bioengineering↗

Simple Feedback for Complex Movement: Capturing Whole-Limb Reorganization during Single-IMU Gait Retraining

Clinical gait retraining typically relies on multi-sensor arrays and high-dimensional feedback displays, imposing setup and interpretation burdens that limit routine clinical deployment. We developed a single-IMU visual biofeedback system that delivers real-time feedback of Lower Limb Trajectory Error (LLTE), a composite kinematic error metric integrating knee position and shank angle across the stance phase. Twenty able-bodied adults walked on a treadmill under two visual biofeedback targets (flexed-knee, extended-knee) while receiving either corrected (n=10) or uncorrected (n=8) feedback, where the correction accounted for limb orientation at initial contact. LLTE and stance-phase knee kinematics adapted consistently under the flexed-knee target for both feedback groups, with feedback formulation moderating the temporal trajectory of change. Adaptation toward the extended-knee target was limited, likely because participants were already operating near terminal knee extension and because the scalar error metric provided limited directional information for correction. Ankle range of motion (ROM) changed significantly across the stance phase under both target conditions, while hip ROM did not. Multiscale multivariate sample entropy (MSMVSE) increased monotonically with time scale across all conditions, with no statistically distinguishable difference between corrected and uncorrected feedback. These results suggest that single-IMU LLTE biofeedback can modify gait mechanics and that adaptation was expressed across multiple lower-limb segments rather than through changes at a single joint.

bioengineering↗