Engineered cells used as smart vehicles for delivery
of secreted
therapeutic proteins enable effective treatment of cancer and certain
degenerative, autoimmune, and genetic disorders. However, current
cell-based therapies use mostly invasive tools for tracking proteins
and do not allow for controlled secretion of therapeutic proteins,
which could result in unconstrained killing of surrounding healthy
tissues or ineffective killing of host cancer cells. Regulating the
expression of therapeutic proteins after success of therapy remains
elusive. In this study, a noninvasive therapeutic approach mediated
by magneto-mechanical actuation (MMA) was developed to remotely regulate
the expression of the tumor necrosis factor-related apoptosis-inducing
ligand (TRAIL) protein, which is secreted by transduced cells. Stem
cells, macrophages, and breast cancer cells were transduced with a
lentiviral vector encoding the SGpL2TR protein. SGpL2TR comprises
TRAIL and GpLuc domains optimized for cell-based applications. Our
approach relies on the remote actuation of cubic-shape highly magnetic
field responsive superparamagnetic iron oxide nanoparticles (SPIONs)
coated with nitrodopamine PEG (ND-PEG), which are internalized within
the cells. Cubic ND-PEG-SPIONs actuated by superlow frequency alternating
current magnetic fields can translate magnetic forces into mechanical
motion and in turn spur mechanosensitive cellular responses. Cubic
ND-PEG-SPIONs were artificially designed to effectively operate at
low magnetic field strengths (<100 mT) retaining approximately
60% of their saturation magnetization. Compared to other cells, stems
cells were more sensitive to the interaction with actuated cubic ND-PEG-SPIONs,
which clustered near the endoplasmic reticulum (ER). Luciferase, ELISA,
and RT-qPCR analyses revealed a marked TRAIL downregulation (secretion
levels were depleted down to 30%) when intracellular particles at
0.100 mg/mL Fe were actuated by magnetic fields (65 mT and 50 Hz for
30 min). Western blot studies indicated actuated, intracellular cubic
ND-PEG-SPIONs can cause mild ER stress at short periods (up to 3 h)
of postmagnetic field treatment thus leading to the unfolded protein
response. We observed that the interaction of TRAIL polypeptides with
ND-PEG can also contribute to this response. To prove the applicability
of our approach, we used glioblastoma cells, which were exposed to
TRAIL secreted from stem cells. We demonstrated that in the absence
of MMA treatment, TRAIL essentially killed glioblastoma cells indiscriminately,
but when treated with MMA, we were able to control the cell killing
rate by adjusting the magnetic doses. This approach can expand the
capabilities of stem cells to serve as smart vehicles for delivery
of therapeutic proteins in a controlled manner without using interfering
and expensive drugs, while retaining their potential to regenerate
damaged tissue after treatment. This approach brings forth new alternatives
to regulate protein expression noninvasively for cell therapy and
other cancer therapies.