Sleeping Beauty transposon system for GDNF overexpression of entrapped stem cells in fibrin hydrogel in a rat model of Parkinson’s disease

Abstract

There is currently no causal treatment available for Parkinson’s disease (PD). However, the use of glial cell line–derived neurotrophic factor (GDNF) to provide regenerative effects for neurons is promising. Such approaches require translational delivery systems that are functional in diseased tissue. To do so, we used a non-viral Sleeping Beauty (SB) transposon system to overexpress GDNF in adipose tissue–derived mesenchymal stromal cells (adMSCs). Entrapment of cells in fibrin hydrogel was used to boost potential neurorestorative effects. Functional GDNF-adMSCs were able to secrete 1066.8 ± 169.4 ng GDNF/120,000 cells in vitro. The GDNF-adMSCs were detectable for up to 1 month after transplantation in a mild 6-hydroxydopamine (6-OHDA) hemiparkinson male rat model. Entrapment of GDNF-adMSCs enabled GDNF secretion in surrounding tissue in a more concentrated manner, also tending to prolong GDNF secretion relatively. GDNF-adMSCs entrapped in hydrogel also led to positive immunomodulatory effects via an 83% reduction of regional IL-1β levels compared to the non-entrapped GDNF-adMSC group after 1 month. Furthermore, GDNF-adMSC-treated groups showed higher recovery of tyrosine hydroxylase (TH)-expressing cells, indicating a neuroprotective function, although this was not strong enough to show significant improvement in motor performance. Our findings establish a promising GDNF treatment system in a PD model. Entrapment of GDNF-adMSCs mediated positive immunomodulatory effects. Although the durability of the hydrogel needs to be extended to unlock its full potential for motor improvements, the neuroprotective effects of GDNF were evident and safe. Further motor behavioral tests and other disease models are necessary to evaluate this treatment option adequately. Graphical Abstract

Advanced Progress in the Role of Adipose-Derived Mesenchymal Stromal/Stem Cells in the Application of Central Nervous System Disorders

“…Utilizing a non-viral Sleeping Beauty (SB) transposon system, Stahn et al engineered GDNF-ADMSCs and introduced them into a mild 6-OHDA hemiparkinson male rat model. The findings highlighted the immunomodulatory and neuroprotective attributes of GDNF-ADMSCs, including the restoration of tyrosine hydroxylase-expressing cells [106] (Table 2).…”

Transposon delivery for CRISPR-based loss-of-function screen in mice identifies NF2 as a cooperating gene involved with the canonical WNT signaling molecular class of hepatocellular carcinoma

“…In previous SB transposon screens, NF2 was identified as a candidate gene for osteosarcoma [ 48 ], while SAV1 was identified in a NAFLD screen and its liver-specific deletion resulted in lipid accumulation, apoptosis and fibrogenesis [ 49 ]. While the SB system was used to overexpress GDNF as potential treatment for Parkinson’s disease [ 50 , 51 ].…”

Uncovering Novel Therapeutic Targets for Parkinson’s Disease

“…This GDNF with MSCs were transplanted in rats treated with 6-OHDA. Up to 1 month, they were detectable, and they were able to provide significant neuroprotective action by increasing TH density and reducing levels of inflammatory cytokines . In another study, GDNF was loaded into lipid nanoparticles and administered intranasally in a MPTP induced mouse model of PD.…”

“…Up to 1 month, they were detectable, and they were able to provide significant neuroprotective action by increasing TH density and reducing levels of inflammatory cytokines. 156 In another study, GDNF was loaded into lipid nanoparticles and administered intranasally in a MPTP induced mouse model of PD. These lipid nanocarriers were coated with chitosan and conjugated with transactivator of transcription (TAT).…”