Preclinical Imaging

Invicro can perform competitive tissue binding assays comparing labelled and non-radioactive drugs using a combination of in vitro assays and high-resolution ex vivo imaging techniques to confirm radioligand binding and inform dose selection for subsequent studies.

Target densities (maximum binding signal, B_max) and regional distributions may not be known. In order to determine these parameters, Invicro can utilize multiple approaches to measure target expression in ethically sourced animal and/or human tissue including:

• in vitro autoradiography
• Homogenate binding assays
• Bioactivity assays
• Internalization Assays
• in vivo Imaging
• FACS
• Western Blotting

Invicro is a leading provider of optical imaging services to determine the efficacy of therapeutic candidates. In this example, efficacy of anti-tumor therapies targeting deep tissue tumor models (orthotopic) was determined non-invasively using Bioluminescence. This imaging approach allowed for efficacy to be determined with tissue relevant context. Invicro has experience working with a wide range of tumor models including:

• Hematological
• Solid Tumors
• Metastatic

Dynamic contrast enhanced (DCE) T1-weighted MRI provides functional characterization of tumor perfusion and permeability, which may be heterogeneous. We have developed analysis tools for semi-automated estimation of parametric maps or ROI analysis.

Invicro has extensive experience running high throughput biodistribution studies in preclinical animal models. This service allows for the following experimental parameters to be determined:

• Whole body biodistribution
• Kinetics
• Off-target accumulation

Invicro has extensive experience labeling different classes of molecules for in vivo and ex vivo imaging applications. From novel radioligand labeling using a risked-managed approach to meet specific customer needs, to GMP-based labeling for clinical imaging, the Invicro Chemistry team has the expertise required to label imaging agents using long and short-lived half life radionuclides for SPECT and PET along with fluorescence for optical imaging applications. We have direct experience with labeling the following types of molecules:

• Small Molecules
• Antibodies, Antibody Fragments
• Cells (T-cells)
• Viruses
• Liposomes
• Exosomes
• Nucleic Acids (ASO, RNA, DNA)
• Novel ligands

Invicro provides single voxel, in vivo 1H MRS. This technique can be used for longitudinal monitoring and therapy response of various metabolites and processes in parallel, for example, neurochemicals (Glu/Gln and GABA) and energy metabolism (PCr, lactate, and ATP). This method is also translatable to clinic. Invicro has various pulse sequences available (PRESS, semiLASER, ISIS, and STEAM) in addition to an automated LC Model (LCModel and Provencher) analysis workflow. Other applications are tissue pH (intra- and extracellular by P-31), quantification, distribution, and metabolism of exogenous F-19 labelled compounds as well as high-resolution ex vivo MR spectroscopy of tissue extracts.

Invicro’s cryo-fluorescence tomography (CFT) is a 3D, high resolution imaging modality that can track the biodistribution of various test articles, including gene and cellular therapies delivered using viruses and lipid nanoparticles. Registration to white light images and fly through movie generation provides accurate localization of signal. Visualization of anatomical details with high soft tissue contrast using in vivo MRI can evaluate morphological changes in brain, tumors, and more and detect small lesions (e.g., in the lung). The spatial resolution can be tailored as needed, as in the example below, for details such as optical nerves. Invicro can employ all standard MR contrasts and has access to various RF coils for whole body or body parts, with gating for improved spatial resolution and customizable analysis. CT and ultrasound are also available for structural imaging.

Combine preclinical discovery work with distributed tumor model simulations to directly guide clinical imaging protocols to support the estimation of tumor properties in human subjects and guide the design of first-in-human oncology clinical imaging protocols with novel biomarkers.

Quantifying changes in BOLD. This allows in vivo, longitudinal assessment of non-labelled compounds. Various mathematical modelling and simulation methods are available. Brain atlases are available for various species, including rodents and non-human primates.

Perceptive uses pharmacokinetic modeling to support study design and interpretation of experimental data. Invicro often provides PK modeling support in an iterative fashion to continually learn and modify the model and/or model parameters to build confidence in simulations to support experimental study design and clinical translation. The figure uses PK modeling to demonstrate the impact of molecular weight on tumor accumulation while keeping all other model parameters constant.

Bringing new therapeutic radiopharmaceuticals to the market is a complex and challenging process. When filing an Investigational New Drug (IND) or Clinical Trial Application (CTA) prior to the initiation of human trials, it is crucial to have strong non-clinical data to support a new candidate. Invicro provides complete preclinical support of your project, including:

• Study design and consultation
• Radiolabeling/conjugation to support alpha, beta, and gamma emitting isotopes
• in vitro/ex vivo cell and tissue-based assays
• Primary pharmacology studies to demonstrate MOA
• in vivo Imaging model development to support theranostic imaging
• Advanced image analytics
• Dosimetry
• IND support
• Development of chemistry, manufacturing and controls package for imaging agents

Bioluminescence imaging (BLI) using luciferase reporters has revolutionized discovery research and is now a mainstay for in vivo testing in oncology (e.g., oncolytic virus and immune cell tracking) and for evaluating gene therapies delivered using lipid nanoparticles, viruses, and siRNA. In this example, different mRNA gene products were evaluated with or without supplemental test agents to determine effectiveness in promoting expression.

Gene delivery agent screening study: in vivo (italics in vivo) mRNA gene delivery time course for multiple test agents (route: IM) quantified by bioluminescence imaging. The graph shows hindlimb photon flux (p/s) vs. time, with the data plotted as geometric mean +/- standard error in geometric mean. Example images highlight a combination treatment benefit vs. Single agent for test Test Agent 2.

Visualization of anatomical details with high soft tissue contrast using in vivo MRI can evaluate morphological changes in brain, tumors, and more and detect small lesions (e.g., in the lung). The spatial resolution can be tailored as needed, as in the example below, for details such as optical nerves. Invicro can employ all standard MR contrasts and has access to various RF coils for whole body or body parts, with gating for improved spatial resolution and customizable analysis. CT and ultrasound are also available for structural imaging.

Diffusion-based MRI can be used to localize brain damage and monitor disease progression monitoring. Modelling for quantitative parametric maps is available.

Obtain segmentation, calculation, and visualization of vessel and radius and density. View a deeper branch-by-branch analysis of branch radius, length, vessel branching fraction, and tortuosity with characterization of vessel properties