Methods and compositions for treating skin disease with recombinant microorganisms
The present invention provides isolated plasmids, recombinant microorganisms, kits, and methods for the treatment of inflammatory skin disease.
1. A recombinant microorganism capable of secreting a filaggrin polypeptide, wherein the recombinant microorganism is Staphylococcus epidermidis , comprising an expression vector comprising a first coding sequence comprising a gene capable of expressing the filaggrin polypeptide and a second coding sequence comprising a gene capable of expressing a cell penetrating peptide, wherein the filaggrin polypeptide comprises an amino acid sequence at least 98% identical to SEQ ID NO: 1.
2. The recombinant microorganism of claim 1 , further comprising a third coding sequence comprising a gene capable of expressing an export signal.
3. The recombinant microorganism of claim 1 , wherein the microorganism secretes a filaggrin fusion protein.
4. A live biotherapeutic composition comprising a recombinant microorganism capable of secreting a filaggrin polypeptide, wherein the recombinant microorganism is Staphylococcus epidermidis , comprising
(i) a first coding sequence comprising a nucleic acid sequence capable of expressing the filaggrin therapeutic polypeptide, wherein the filaggrin polypeptide comprises an amino acid sequence at least 98% identical to SEQ ID NO: 1;
(ii) a second coding sequence comprising a nucleic acid sequence capable of expressing a cell penetrating peptide;
(iii) a third coding sequence comprising a nucleic acid sequence capable of expressing an export signal; and
(iv) a promoter operably linked to the first coding sequence, the second coding sequence and the third coding sequence;
wherein the first coding sequence, second coding sequence and first coding sequence is capable of expressing a filaggrin fusion product, or variant thereof.
5. The composition of claim 4 , comprising a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier is selected from the group consisting of an aqueous solution, an emulsion, a cream, a lotion, a gel, or an ointment.
6. A method for producing a live biotherapeutic composition, the method comprising:
(a) transfecting a Staphylococcus epidermidis cell with an expression vector comprising a first coding sequence comprising a gene capable of expressing the filaggrin polypeptide and a second coding sequence comprising a gene capable of expressing a cell penetrating peptide, wherein the filaggrin polypeptide comprises an amino acid sequence at last 98% identical to SEQ ID NO: 1; and
(b) allowing the transfected cell to produce a therapeutic polypeptide fusion protein; and
(c) obtaining the live biotherapeutic composition.
7. The method of claim 6 , further comprising (iii) transfecting the cell with a third coding sequence comprising a nucleic acid sequence capable of expressing an export signal.
8. A method of treating a skin disease comprising administering to a subject in need thereof the composition of claim 1 .
9. The method of claim 8 , wherein the skin disease is Ichthyosis vulgaris (IV).
10. The method of claim 8 , wherein the skin disease is atopic dermatitis.