Human Cord Blood-Derived Unrestricted Somatic Stem Cell Infusion Improves Neurobehavioral Outcome in a Rabbit Model of Intraventricular Hemorrhage

Abstract

Intraventricular hemorrhage (IVH) is a severe complication of preterm birth, which leads to hydrocephalus, cerebral palsy, and mental retardation. There are no available therapies to cure IVH, and standard treatment is supportive care. Unrestricted somatic stem cells (USSCs) from human cord blood have reparative effects in animal models of brain and spinal cord injuries. USSCs were administered to premature rabbit pups with IVH and their effects on white matter integrity and neurobehavioral performance were evaluated. USSCs were injected either via intracerebroventricular (ICV) or via intravenous (IV) routes in 3 days premature (term 32d) rabbit pups, 24 hours after glycerol‐induced IVH. The pups were sacrificed at postnatal days 3, 7, and 14 and effects were compared to glycerol‐treated but unaffected or nontreated control. Using in vivo live bioluminescence imaging and immunohistochemical analysis, injected cells were found in the injured parenchyma on day 3 when using the IV route compared to ICV where cells were found adjacent to the ventricle wall forming aggregates; we did not observe any adverse events from either route of administration. The injected USSCs were functionally associated with attenuated microglial infiltration, less apoptotic cell death, fewer reactive astrocytes, and diminished levels of key inflammatory cytokines (TNFα and IL1β). In addition, we observed better preservation of myelin fibers, increased myelin gene expression, and altered reactive astrocyte distribution in treated animals, and this was associated with improved locomotor function. Overall, our findings support the possibility that USSCs exert anti‐inflammatory effects in the injured brain mitigating many detrimental consequences associated with IVH. stem cells translational medicine 2019;8:1157–1169

Human Cord Blood Derived Unrestricted Somatic Stem Cells Restore Aquaporin Channel Expression, Reduce Inflammation and Inhibit the Development of Hydrocephalus After Experimentally Induced Perinatal Intraventricular Hemorrhage

“…On the basis of USSC functions and our prior report (Vinukonda et al, 2019 ), we hypothesized that USSC administration would stabilizes the aquaporin water channels in the choroid plexus and ependymal wall resulting in reduced hydrocephalus after IVH. Further, we hypothesized that USSC administration would also suppress inflammation in the choroid plexus (TLR2, TLR4 and NF-kB) as well as ventricular wall matrix reorganization (TGF-β isoforms, matrix metalloproteinase-9, MMP-9), and promote anti-inflammatory cytokine (Interleukin-10; IL-10) expression which collectively would contribute to the attenuation of PhH after IVH.…”

“…USSCs were isolated from human umbilical cord blood mononuclear cells according to the methods of Kogler and colleagues as we have previously described (Kogler et al, 2004 ; Liao et al, 2014 ; Vinukonda et al, 2019 ). USSCs were transduced with a lentivirus carrying GFP-luciferase gene prepared using the lenti-viral construct, pSico PolII-eGFP-Luc2 (generously provided by Dr. Glenn Merlino at the National Cancer Institute) to enable in vivo identification while enabling the cells to retain their functionality (Liao et al, 2014 ).…”

“…These reports showed improved myelination, neuroprotection and less apoptosis (Liu et al, 2010 ; Ahn et al, 2014 ; Mukai et al, 2017 ). To expand upon these observations, our group investigated unrestricted somatic stem cells (USSCs) from human cord blood that were engineered to express the luciferase reporter gene, confirmed a stable non-teratogenic phenotype and successfully tracked USSC in vivo migration in a rabbit model of intraventricular hemorrhage IVH (Vinukonda et al, 2019 ). We selected USSCs because they release growth factors and cytokines with known neuroprotective and axonal growth promoting functions at higher levels than MSCs and for their anti-inflammatory and immunomodulatory properties.…”

Retrieval of germinal zone neural stem cells from the cerebrospinal fluid of premature infants with intraventricular hemorrhage

“…Our data partly support those studies, although, unlike Kruegger et al, we were not able to isolate NSCs from nonhemorrhagic CSF samples and it is likely that cells obtained by these authors from indwelling ventricular catheters represent a different population. Currently, stem cell therapies based on allogeneic umbilical cord‐derived cells are being tested in preclinical and clinical trials 63‐67 . These cells help to attenuate the effects associated to inflammation but, so far, no capability to replace lost tissue has been demonstrated.…”

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“…Meanwhile, UCB‐MSCs display similarities to MSCs derived from other tissues and, hence, may find use in therapies for a wide range of immune or inflammatory disorders such as atopic dermatitis. In our second Featured Article this month published in Stem Cells Translational Medicine , Vinukonda et al report that cord blood‐derived USSCs administered in a rabbit model of intraventricular hemorrhage (IVH) decreased microglial infiltration, pro‐inflammatory cytokine expression, and apoptotic cell loss, thereby prompting the partial recovery of myelination, preservation of white matter, and recovery of motor function . In a Related Article published in Stem Cells , Kim et al established the efficacy and safety of a single subcutaneous injection of human (h)UCB‐MSCs in patients with moderate‐to‐severe atopic dermatitis .…”