Human Gene Therapy - April 2023 - 311

EX VIVO GENE DELIVERY TO PORCINE HEARTS USING AAV
311
Figure 6. Anti-luciferase immunofluorescent staining of the allograft versus native cardiac tissues. Robust and homogeneous luciferase distribution was
observed in the cardiac allograft treated with (A) 8 · 1013 VGC, whereas (B) no luciferase was detected in the recipient's native heart. Similarly, there was
robust and homogeneous luciferase distribution observed in the cardiac allograft treated with (C) 1 · 1014 VGC, but (D) no luciferase detected in the recipient's
native heart.
periments describing successful gene delivery to rabbit
donor hearts were reported by Shah et al.26 They utilized
recombinant Ad vectors delivered ex vivo to hearts into
the aortic root during a period of cold static preservation.
While global functional changes were described in the
transplanted hearts that were transduced with badrenergic
receptor genes, transgene expression was
noted to be heterogeneous in the left and right ventricles.
Following the growing utility of EVMP in clinical applications,
our group reported that homogeneous and
robust transgene expression was feasible using Ad vectors
delivered through normothermic ex vivo perfusion
using the OCS device.7 While this earlier work was
seminal for establishing effective gene delivery techniques
that can be applied during cardiac transplantation,
these studies were limited in that they conducted their
assessments at 5 days posttransduction, which does not
support widespread clinical application. For this reason,
a longer term and presumably more homogeneous expression
would be required.
The current study builds on these early observations;
delivery of rAAV vector is accomplished during a period
of EVMP to better achieve homogeneous exposure of the
allograft to viral vector and sustain durable transgene expression
posttransplantation.
Using a cardiac-enhanced rAAV, SASTG, we were
able to confer durable transgene expression after a 2-h
transduction period during EVMP. We approached our
choice of AAV capsid based on our in vitro experiments
outlined in this study showing that SASTG was the most
efficient of those we tested at delivering transgenes and
that its performance is comparable from in vitro to in vivo
applications. Our analysis suggests that the majority of
vector uptake by the allograft or the circuit occurs within
the first 30 min. We observed diffuse transgene expression
throughout the graft at each of the three escalating vector
doses. This is the first demonstration that a rAAV delivered
during EVMP can confer transgene expression in a
robust, global manner in a large-animal allograft, with no
off-target gene expression or organ injury.

Human Gene Therapy - April 2023

Table of Contents for the Digital Edition of Human Gene Therapy - April 2023

Contents
Human Gene Therapy - April 2023 - CT1
Human Gene Therapy - April 2023 - CT2
Human Gene Therapy - April 2023 - Cover1
Human Gene Therapy - April 2023 - Cover2
Human Gene Therapy - April 2023 - 239
Human Gene Therapy - April 2023 - 240
Human Gene Therapy - April 2023 - 241
Human Gene Therapy - April 2023 - 242
Human Gene Therapy - April 2023 - 243
Human Gene Therapy - April 2023 - 244
Human Gene Therapy - April 2023 - Contents
Human Gene Therapy - April 2023 - 246
Human Gene Therapy - April 2023 - 247
Human Gene Therapy - April 2023 - 248
Human Gene Therapy - April 2023 - 249
Human Gene Therapy - April 2023 - 250
Human Gene Therapy - April 2023 - 251
Human Gene Therapy - April 2023 - 252
Human Gene Therapy - April 2023 - 253
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Human Gene Therapy - April 2023 - 257
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Human Gene Therapy - April 2023 - 261
Human Gene Therapy - April 2023 - 262
Human Gene Therapy - April 2023 - 263
Human Gene Therapy - April 2023 - 264
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Human Gene Therapy - April 2023 - 268
Human Gene Therapy - April 2023 - 269
Human Gene Therapy - April 2023 - 270
Human Gene Therapy - April 2023 - 271
Human Gene Therapy - April 2023 - 272
Human Gene Therapy - April 2023 - 273
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Human Gene Therapy - April 2023 - 276
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Human Gene Therapy - April 2023 - 310
Human Gene Therapy - April 2023 - 311
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Human Gene Therapy - April 2023 - Cover3
Human Gene Therapy - April 2023 - Cover4
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