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 microorganism is selected from the group consisting of Bifidobacterium, Brevibacterium, Propionibacterium, Lactococcus, Streptococcus, Staphylococcus, Lactobacillus, Enterococcus, Pediococcus, Leuconostoc , or Oenococcus , wherein the recombinant microorganism comprises an expression vector comprising a first coding sequence comprising a gene capable of expressing the 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 95% identical to SEQ ID NO:15.
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 recombinant microorganism is Staphylococcus epidermidis.
4 . The recombinant microorganism of claim 1 , wherein the microorganism secretes a filaggrin fusion protein.
5 . A method for producing a live biotherapeutic composition, the method comprising:
(a) transfecting a cell with (i) a first coding sequence comprising a nucleic acid sequence capable of expressing a filaggrin polypeptide, and (ii) a second coding sequence comprising a nucleic acid sequence capable of expressing a cell penetrating peptide, wherein the cell is a bacterial cell selected from the group consisting of Bifidobacterium, Brevibacterium, Propionibacterium, Lactococcus, Streptococcus, Staphylococcus, Lactobacillus, Enterococcus, Pediococcus, Leuconostoc , or Oenococcus , wherein the filaggrin polypeptide comprises an amino acid sequence at least 95% identical to SEQ ID NO: 15; and
(b) allowing the transfected cell to produce a therapeutic polypeptide fusion protein; and
(c) obtaining the live biotherapeutic composition.
6 . The method of claim 5 , further comprising (iii) transfecting the cell with a third coding sequence comprising a nucleic acid sequence capable of expressing an export signal.
7 . A composition obtained by the method of claim 5 .
8 . The composition of claim 7 , 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.
9 . A live biotherapeutic composition comprising a recombinant microorganism wherein the recombinant microorganism comprises
(i) a first coding sequence comprising a nucleic acid sequence capable of expressing a therapeutic polypeptide;
(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.
10 . The composition of claim 9 , wherein the recombinant microorganism is Staphylococcus epidermidis.
11 . A method of treating a skin disease comprising administering to a subject in need thereof the composition of claim 1 .
12 . The method of claim 11 , wherein the skin disease is Ichthyosis vulgaris (IV).
13 . The method of claim 11 , wherein the skin disease is atopic dermatitis.