Toward Engineered Biosynthesis of Drugs in Human Cells.

Area of Science:

• Biochemistry
• Molecular Biology
• Synthetic Biology

Background:

• Biosynthetic genes are crucial for producing bioactive substances.
• Cytochrome P450 enzymes are key in natural product bioactivity.
• Heterologous gene expression offers potential for novel applications like gene therapy.

Purpose of the Study:

• To assess the feasibility of human cells producing antibiotics in situ.
• To investigate the use of a heterologous biosynthetic gene (Fma-P450) in human cells (HeLa).
• To explore the potential of gene therapy applications for antibiotic production.

Main Methods:

• HeLa cells were engineered to express the fungal Fma-P450 gene.
• Cells were supplemented with the precursor β-trans-bergamotene.
• Production of fungal metabolites by the engineered HeLa cells was analyzed.

Main Results:

• HeLa cells expressing Fma-P450 successfully biosynthesized 5-hydroxyl-β-trans-bergamoten.
• The cytotoxic compound 5-epi-demethoxyfumagillol was also produced.
• Antibiotic production levels were insufficient to cause cell death.

Conclusions:

• Programming human cells to autogenerate antibiotics via heterologous gene introduction is feasible.
• This study demonstrates a proof-of-concept for in situ antibiotic production in human cells.
• Further research may optimize production levels for therapeutic applications.