Enhancement of placental stem cell potency using modulatory RNA molecules

• US 8,969,315 B2
• Filed: 12/29/2011
• Issued: 03/03/2015
• Est. Priority Date: 12/31/2010
• Status: Active Grant

• Current Assignee: Celularity, Inc.
• Original Assignee: Anthrogenesis Corporation (Bristol-Myers Squibb Company)
• Inventors: Abbot, Stewart, Voskinarian-Berse, Vanessa, Zhang, Xiaokui, Karasiewicz-Mendez, Kathy E.
• Primary Examiner(s): Chong, Kimberly
• Application Number:
  US13/340,589
• Publication Number:
  US 20120230959A1
• Time in Patent Office: 1,160 Days
• Field of Search: None
• US Class Current: 514/44
• CPC Class Codes: A61K 2035/122 for inducing tolerance or supression of immune responses
• CPC Class Codes: A61K 31/7088 Compounds having three or more nucleosides or nucleotides
• CPC Class Codes: A61K 31/7105 Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links
• CPC Class Codes: A61K 31/713 Double-stranded nucleic acids or oligonucleotides
• CPC Class Codes: A61K 35/50 Placenta; Placental stem cells; Amniotic fluid; Amnion; Amniotic stem cells
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: C12N 15/1138 against receptors or cell surface proteins
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/141 MicroRNAs, miRNAs
• CPC Class Codes: C12N 2310/531 Stem-loop; Hairpin
• CPC Class Codes: C12N 2501/38 with nuclear receptors
• CPC Class Codes: C12N 2501/65 MicroRNA
• CPC Class Codes: C12N 5/0605 Cells from extra-embryonic tissues, e.g. placenta, amnion, yolk sac, Wharton's jelly
• CPC Class Codes: C12N 5/0668 Mesenchymal stem cells from other natural sources

Abstract

Provided herein are methods of producing enhanced placental stem cells by modulatory RNA molecules. Also provided herein are methods of using enhanced placental stem cells, for example, to treat individuals having a disease, disorder or condition caused by, or relating to, an unwanted or harmful immune response. Further provided herein are compositions comprising said enhanced placental stem cells.

Claims

1. A method of treating an individual having or at risk of developing a disease, disorder or condition having an inflammatory component, comprising administering to the individual a therapeutically effective amount of enhanced placental stem cells,

wherein said enhanced placental stem cells comprise or have been contacted with an effective amount of modulatory RNA molecules that decrease expression of one or more human nuclear receptors in said enhanced placental stem cells, resulting in suppression of soluble IL-23 protein produced by peripheral blood mononuclear cells (PBMCs) in the presence of said enhanced placental stem cells compared to placental stem cells not contacted with said modulatory RNA molecules,

wherein said modulatory RNA molecules are small interfering RNAs (siRNAs), microRNA inhibitors (miR inhibitors), or micro RNA mimics (miR mimics),

wherein said one or more human nuclear receptors is selected from the group consisting of:

vitamin D receptor (VDR);

nuclear receptor subfamily 4, group A, member 3 (NR4A3);

nuclear receptor subfamily 0, group B, member 2 (NR0B2);

nuclear receptor subfamily 1, group 1, member 2 (NR1I2);

nuclear receptor subfamily 1, group H, member 3 (NR1H3); and

deoxynucleotidyltransferase, terminal, interacting protein 1 (DNTTIP1), and

wherein the therapeutically effective amount is an amount sufficient to cause a detectable improvement in one or more symptoms of said disease, disorder or condition.

2. The method of claim 1, wherein said modulatory RNA molecules are siRNAs.

3. The method of claim 1, wherein said modulatory RNA molecules are selected from a library.

4. The method of claim 3, wherein said library is a human nuclear receptor library, human phosphatase siRNA library, or an anti-miR library.

5. The method of claim 1, wherein said placental stem cells are CD10⁻

, CD34⁻

, CD105⁺, CD200⁺ placental stem cells.

6. The method of claim 1, wherein said placental stem cells express CD200 and do not express HLA-G, or express CD73, CD105, and CD200, or express CD200 and OCT-4, or express CD73 and CD105 and do not express HLA-G.

7. The method of claim 5, wherein said placental stem cells are additionally CD90⁺ and CD45⁻

.

8. The method of claim 5, wherein said placental stem cells are additionally CD80⁻

and CD86⁻

.