Neural Derivates of Canine Induced Pluripotent Stem Cells-Like Cells From a Mild Cognitive Impairment Dog

Abstract

Domestic dogs are superior models for translational medicine due to greater anatomical and physiological similarities with humans than rodents, including hereditary diseases with human equivalents. Particularly with respect to neurodegenerative medicine, dogs can serve as a natural, more relevant model of human disease compared to transgenic rodents. Herein we report attempts to develop a canine-derived in vitro model for neurodegenerative diseases through the generation of induced pluripotent stem cells from a 14-year, 9-month-old female West Highland white terrier with mild cognitive impairment (MCI). Canine induced pluripotent stem cells-like cells (ciPSCLC) were generated using human OSKM and characterized by positive expression of pluripotency markers. Due to inefficient viral vector silencing we refer to them as ciPSCLCs. Subsequently, the ciPSCLC were subjected to neural induction according to two protocols both yielding canine neural progenitor cells (cNPCs), which expressed typical NPC markers. The cNPCs were cultured in neuron differentiation media for 3 weeks, resulting in the derivation of morphologically impaired neurons as compared to iPSC-derived human counterparts generated in parallel. The apparent differences encountered in this study regarding the neural differentiation potential of ciPSCLC reveals challenges and new perspectives to consider before using the canine model in translational neurological studies.

Induced pluripotent stem cells in companion animals: how can we move the field forward?

“…While colonies could be obtained with this method it was only when highly defined media was used, and still the reprogramming efficiency was very low ( 94 ). Chandrasekaran et al also used episomal reprogramming with electroporation, but even though morphological changes were observed in the cells, complete reprograming was not achieved and lentivirus was used subsequently, resulting in generation of canine iPSCs from the same somatic cells ( 95 ).…”

“…When it comes to the applicability of iPSCs in companion animals, reports are scarcer than for their generation and mainly focus on therapeutics or disease modeling. Therapeutic use of iPSCs has been mainly proposed for musculoskeletal conditions in horses ( 60 – 63 , 88 , 127 – 129 ) and for neurological and cardiovascular conditions in dogs ( 67 , 95 , 130 – 133 ). In terms of disease modeling, equine iPSCs can also be used for neurological conditions ( 58 , 134 ).…”

“…Compared to current advances in the use of human iPSCs in disease modeling and drug screening, there is scarce published literature in companion animals and is mostly limited to neurological disorders. In canines, an iPSCs line was generated from a West Highland White Terrier affected by mild cognitive impairment, showing an important proof of concept on successfully generating iPSCs from a geriatric patient ( 95 ), while on the equine side iPSC-based in vitro models have been generated to study the process of neurotropic viral infections ( 134 ). Although limited, these studies constitute relevant milestones for generating efficient in vitro modeling systems that will not depend on limitations of adult somatic cells and can eventually lead to a personalized/customized medicine approach where the most efficient treatment will be made available in a patient-centered approach.…”

Opening the “Black Box” Underlying Barriers to the Use of Canine Induced Pluripotent Stem Cells: A Narrative Review

“…While this canine model demonstrated successful delivery, undifferentiated ciPSCs are not directly amenable for clinical transplantation due to higher risk of teratoma formation [ 22 ]. ciPSCs can be differentiated into either progenitor cells or adult cells such as neural, cardiac, endothelial, and platelets [ 16 , 22 , 38 , 93–96 ]. A summary of research studying the clinical applications of ciPSCs to date is shown in Table 3 .…”

“…A similar study to Chow et al (2020) by Chandrasekaran et al (2021) differentiated ciPSC-like cells into NPCs that were used to repair damaged nervous tissue in dogs [ 93 ]. They found that despite incomplete reprogramming of ciPSCs, canine NPCs had reduced axonal fasciculation and shorter neuritis, demonstrating lower mature neuronal differentiation efficiency than human NPCs [ 93 ].…”