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Design of trans-splicing RNA

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Researchers
Volker Patzel
Managed By
Dr Ler Ser Yeng
Manager (65)66016248
Patent Protection

PCT Patent Application WO2017/171654
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MARKET OPPORTUNITY

Gene therapy is considered to be a highly effective treatment for chronic diseases with minimum side effects. The global cancer gene therapy market was valued at $289 million in 2016, and is estimated to reach $2,082 million by 2023, at a compound annual growth rate of 32.4% from 2017 to 2023.

TECHNOLOGY

An advanced tsRNA design towards a Herpes simplex virus thymidine kinase/ganciclovir (HSV-tk/GCV) suicide gene therapy approach. Tested for hepatocellular carcinoma and cervical cancer.


Figure 1: Design of tsRNA for 5′ and 3′ exon replacement (ER) and structure of an alpha fetoprotein (AFP) mini-gene.

Molecular targets:

1. Hepatocellular carcinoma (HCC): alpha-fetoprotein (AFP), hepatocellular carcinoma-associated gene 2/ Yin Yang 1-associated protein 1 (HCCA2/YY1AP1), cluster of differentiation 24 (CD24) and Vascular endothelial growth factor (VEGF)

2. Cervical cancer: HPV-16

In vitro anti-tumour efficacy:

1. Hepatocellular carcinoma: 

a. Investigation of death of HepG2 cells triggered by 5’ or 3’ exon replacement (ER) toward overexpressed and endogenous AFP pre-mRNA using AlamarBlue cell viability assay:

i. up to 80% at 100 μM or 60% to 70% at 10 μM GCV

ii. No cytotoxicity was triggered by the AFP mini-gene or the trans-splicing constructs in the absence of GCV.

b. Investigation of death of HepG2 cells triggered by 5’ or 3’ ER toward overexpressed and endogenous AFP pre-mRNA and another HCC biomarker (HCCA2/YY1AP1, CD24, or VEGF as secondary targets) using AlamarBlue cell viability assay:

i. At 10 μM GCV, all dual-targeting constructs triggered significantly higher levels of cell death compared with the construct targeting AFP only.

ii. At 100 μM GCV, single- and dual-targeting constructs showed comparable effects

2. Cervical cancer:

a. Investigation of death of SiHa and C3 cells triggered by 5’ or 3’ ER toward overexpressed and endogenous HPV-16 pre-mRNA using AlamarBlue cell viability assay:

i. In SiHa cells, all tsRNAs triggered cell death to the same extent as the positive control

ii. In C3 cells, 3’ ER was more efficient than 5’ ER.

CATEGORY

Therapeutics

STAGE OF DEVELOPMENT

TRL3

APPLICATIONS

• Gene therapy

ADVANTAGES

• High efficacy

• Targeted

• Minimum side effects

STATUS

Patent pending.