Lentiviral Vector-mediated Gene Therapy of Hepatocytes <em>Ex Vivo</em> for Autologous Transplantation in Swine

Robert A. Kaiser; Shennen A. Mao; Jaime Glorioso; Bruce Amiot; Clara T. Nicolas; Kari L. Allen; Zeji Du; Caitlin J VanLith; Raymond D. Hickey; Scott L. Nyberg; Joseph B. Lillegard

doi:10.3791/58399

Lentiviral Vector-mediated Gene Therapy of Hepatocytes Ex Vivo for Autologous Transplant...

JoVE Journal
Medicine

A subscription to JoVE is required to view this content. Sign in or start your free trial.

JoVE Journal Medicine

Lentiviral Vector-mediated Gene Therapy of Hepatocytes Ex Vivo for Autologous Transplantation in Swine

Lentiviral Vector-mediated Gene Therapy of Hepatocytes Ex Vivo for Autologous Transplantation in Swine

Full Text

8,590 Views

09:54 min

November 4, 2018

DOI: 10.3791/58399-v

Robert A. Kaiser^1,2, Shennen A. Mao¹, Jaime Glorioso³, Bruce Amiot¹, Clara T. Nicolas¹, Kari L. Allen¹, Zeji Du¹, Caitlin J VanLith¹, Raymond D. Hickey¹, Scott L. Nyberg¹, Joseph B. Lillegard^1,2,4

¹Department of Surgery,Mayo Clinic, ²Midwest Fetal Care Center,Children's Hospitals and Clinics of Minnesota, ³Department of Surgery,Johns Hopkins, ⁴Pediatric Surgical Associates

Overview

This protocol describes ex vivo gene therapy and autologous cell transplantation for treating metabolic diseases, specifically hereditary tyrosinemia type 1. It highlights the advantages of this approach, including reduced immunosuppression and the potential for broader application to various liver diseases.

Key Study Components

Area of Science

Gene therapy
Cell transplantation
Metabolic diseases

Background

Ex vivo gene therapy offers an alternative treatment for inborn errors of metabolism.
This method allows for the assessment of necessary biomass for therapeutic outcomes.
It minimizes the risks associated with immunosuppression.
The technique is adaptable for multiple liver diseases.

Purpose of Study

To treat hereditary tyrosinemia type 1 using autologous cell transplantation.
To establish a platform for addressing various liver diseases.
To visualize and optimize the efficiency of hepatocyte perfusion.

Methods Used

Isolation of porcine hepatocytes.
Ex vivo gene delivery techniques.
Assessment of perfusion efficiency.
Application of autologous cell transplantation.

Main Results

Successful isolation of viable hepatocytes for transplantation.
Demonstrated potential for treating metabolic disorders.
Visualized perfusion efficiency impacts therapeutic outcomes.
Established a foundation for future liver disease treatments.

Conclusions

Ex vivo gene therapy is a promising alternative for metabolic disease treatment.
Autologous cell transplantation can minimize immunosuppression risks.
This approach has the potential for broader applications in liver disease therapy.

Frequently Asked Questions

What is ex vivo gene therapy?

Ex vivo gene therapy involves modifying cells outside the body and then transplanting them back to treat diseases.

How does autologous cell transplantation work?

Autologous cell transplantation uses a patient's own cells to reduce the risk of immune rejection.

What diseases can be treated with this approach?

This method is primarily aimed at metabolic diseases, including hereditary tyrosinemia type 1.

What are the benefits of this therapy?

Benefits include reduced immunosuppression and the ability to treat multiple liver diseases.

Why is perfusion efficiency important?

Perfusion efficiency is crucial for ensuring the viability of hepatocytes during transplantation.

Can this method be applied to other liver diseases?

Yes, the approach is designed to be adaptable for various liver diseases beyond hereditary tyrosinemia.

This protocol is intended to describe porcine hepatocyte isolation and ex vivo gene delivery to cure models of metabolic diseases via autologous cell transplantation. Although this particular model enjoys unique advantages that favor successful therapy, the application is a relevant foundation to address additional diseases and indications.

Ex vivo gene therapy and autologous cell transplantation for the treatment of inborn errors of metabolism in the liver is an important alternative to the therapy for these disorders. And allows us to not only treat these disorders, but figure out the biomass necessary to be able to achieve a therapeutic result. This approach also avoids the significant consequences of immunosuppression and other consequences with different types of therapies.

In the current context, we're using this approach to treat hereditary tyrosinemia type 1. However, the idea is that this will be a platform approach to treat a multitude of different liver diseases with minimal alteration. It's important to be able to visualize this technique because there's some variability when it comes to visualizing the efficiency of the perfusion, which is essential to getting viable hepatocytes for the transplantation.

View the full transcript and gain access to thousands of scientific videos