Genetically modified
Methods and compositions for targeted delivery of biotherapeutics are provided. The compositions comprise bile-sensitive St. thermophilus bacteria modified to release a biotherapeutic agent following bile exposure. Biotherapeutic agents released by the St. thermophilus bacteria disclosed herein include AQ and AQR rich peptides. Methods of the invention comprise administering to a subject a St. thermophilus bacterium modified to release a biotherapeutic agent following bile exposure. Administration of the St. thermophilus bacterium promotes a desired therapeutic response. The bacterium may be modified to express and release AQ or AQR rich peptides which subsequently inhibit cellular apoptosis or reduce mucosal damage. Thus, methods of the invention find use in treating or preventing a variety of gastrointestinal disorders including C. difficile infection and antibiotic-associated diarrhea.
1. A Streptococcus thermophilus bacterium that is genetically modified to express a heterologous polypeptide comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence as set forth in SEQ ID NO: 2 or 8, wherein said heterologous polypeptide is released from said bacterium following exposure to bile.
2. The bacterium of claim 1 , wherein said heterologous polypeptide is a therapeutic polypeptide.
3. The bacterium of claim 1 , wherein said heterologous peptide comprises
an amino acid sequence selected from the group consisting of: SEQ ID NO: 2 and 8.
4. The bacterium of claim 1 , wherein said bacterium does not survive said bile exposure.
5. The bacterium of claim 1 , wherein said Streptococcus thermophilus is selected from the group consisting of:
a) Streptococcus thermophilus NCK2071, having been deposited under ATCC Accession Number PTA-11889; and
b) a probiotic Streptococcus thermophilus bacterium.
6. A method of making a Streptococcus thermophilus bacterium, said method comprising genetically modifying said bacterium to express a heterologous polypeptide comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence as set forth in SEQ ID NO: 2 or 8, wherein said polypeptide is released from said bacterium following exposure to bile.
7. The method of claim 6 , wherein said heterologous polypeptide comprises an amino acid sequence selected from the group consisting of:
SEQ ID NO: 2 and 8.
8. A pharmaceutical composition comprising the bacterium according to claim 1 .