Enhancement of placental stem cell potency using modulatory rna molecules

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.

What is claimed: 
     
         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 cells comprise or have been contacted with an effective amount of modulatory RNA molecules that   (i) suppress an amount of soluble IL-23 protein produced by peripheral blood mononuclear cells (PBMCs) in the presence of said enhanced placental stem cells;   (ii) increase cyclooxygenase II (Cox-2) activity in said enhanced placental stem cells;   (iii) increase an amount of PGE2 produced by said enhanced placental stem cells; or   (iv) reduce the level a pro-inflammatory cytokine produced by enhanced placental stem cells,   compared to placental stem cells not contacted with said modulatory RNA molecules,   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 small interfering RNAs (siRNAs), microRNA inhibitors (miR inhibitors), micro RNA mimics (miR mimics), other modulatory RNA molecules, or any combinations thereof. 
     
     
         3 . The method of  claim 1 , wherein said modulatory RNA molecules suppress an amount of soluble IL-23 protein produced by PBMCs in the presence of said enhanced placental stem cells, compared to placental stem cells not contacted with said modulatory RNA molecules. 
     
     
         4 . The method of  claim 3 , wherein said modulatory RNA molecules are siRNAs. 
     
     
         5 . The method of  claim 4 , wherein said siRNAs modulate one or more of Twinfilin-1, vitamin D receptor tVDR), 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 (NR112), nuclear receptor subfamily 1, group H, member 3 (NR1H3) and deoxynucleotidyltransferase, terminal, interacting protein 1 (DNTT1P1) in said enhanced placental stem cells. 
     
     
         6 . The method of  claim 1 , wherein said modulatory RNA molecules increase an amount of PGE2 produced by said enhanced placental stem cells compared to placental stem cells not contacted with said modulatory RNA molecules. 
     
     
         7 . The method of  claim 6 , wherein said modulatory RNA molecules are miR inhibitors or miRNA mimics. 
     
     
         8 . The method of  claim 7 , wherein said miR inhibitors or miRNA mimics modulate one or more of cholinergic receptor, nicotinic beta 1 (muscle) (CHRNB1), chloride channel 6 (CLCN6), chloride intracellular channel 4 (CLIC4), casein kinase 1, gamma 3 (CSNK1G3), casein kinase 2, alpha prime polypeptide (CSNK2A2), dual specificity phosphatase 1 (DUSP1), potassium channel modulatory factor 1 (KCMF1), potassium voltage-gated channel, shaker-related subfamily, member 3 (KCNA3), potassium inwardly-rectifying channel, subfamily J, member 14 (KCNJ14), potassium voltage-gated channel, delayed-rectifier, subfamily S, member 3 (KCNS3), potassium channel tetramerisation domain containing 13 (KCTD13), hepatocyte growth factor hepapoietin A; scatter factor) (HGF), nuclear receptor subfamily 2, group C, member 2 (NR2C2), phosphodiesterase 1B, calmodulin-dependent (PDE1B), phosphodiesterase 7B (PDE7B), phosphatidylinositol 4-kinase type 2 beta (PI4K2B), phosphoinositide-3-kinase, regulatory subunit 1 (alpha) (PIK3R1), phospholipase C, eta 2 (PLCH2), protein phosphatase, Mg2 + /Mn2 +  dependent, 1D (PPM1D), protein phosphatase, Mg2 + /Mn2 +  dependent, 1G (PPM1G), protein phosphatase 1, regulatory (inhibitor) subunit 2 pseudogene 9 (PPP1R2P9), protein phosphatase 1, regulatory (inhibitor) subunit 3B (PPP1R3B), protein phosphatase 1, regulatory (inhibitor) subunit 9B (PPP1R9B), protein phosphatase 2, catalytic subunit, beta isozyme (PPP2CB), protein tyrosine phosphatase type IVA, member 1 (PTP4A1), protein tyrosine phosphatase, receptor type, K (PTPRK), regulator of G-protein signaling 4 (RGS4), regulator of G-protein signaling 7 binding protein (RGS7BP), regulator of G-protein signaling 8 (RGS8), solute carrier family 16, member 3 (monocarboxylic acid transporter 4) (SLC16A3), solute carrier family 30 (zinc transporter), member 1 (SLC30A1), solute carrier family 35, member A4 (SLC35A4), solute carrier family 38, member 7 (SLC38A7), solute carrier family 41, member 1 (SLC41A1 (includes EG:254428)), solute carrier family 45, member 3 (SLC45A3), solute carrier family 7 (cationic amino acid transporter, y+ system), member 1 (SLC7A1), ubiquitin associated protein 2 (UBAP2), ubiquitin-conjugating enzyme E2D 3 (UBC4/5 homolog, yeast) (UBE2D3 (includes EG:7323)), ubiquitin-conjugating enzyme E2E 3 (UBC4/5 homolog, yeast) (UBE2E3), ubiquitin-conjugating enzyme E2R 2 (UBE2R2 (includes EG:54926)), ubiquitin-conjugating enzyme E2W (putative) (UBE2W), ubiquitin-like with PHD and ring finger domains 2 (UHRF2), ubiquitin specific peptidase 9, X-linked (LUSP9X), and hypoxia inducible factor 1, alpha subunit (HIF1A) in said enhanced placental stem cells. 
     
     
         9 . The method of  claim 1 , wherein said modulatory RNA molecules increase Cox-2 activity in said enhanced placental stem cells compared to placental stem cells not contacted with said modulatory RNA molecules. 
     
     
         10 . The method of  claim 1 , wherein said modulatory RNA molecules reduce the level a pro-inflammatory cytokine produced by enhanced placental stem cells compared to placental stem cells not contacted with said modulatory RNA molecules. 
     
     
         11 . The method of  claim 10 , wherein said pro-inflammatory cytokine is IL-6, IL-8, or a combination thereof. 
     
     
         12 . The method of  claim 1 , wherein said modulatory RNA molecules are selected from a library. 
     
     
         13 . The method of  claim 12 , wherein said library is a human nuclear receptor library, human phosphatase siRNA library, or an anti-miR library. 
     
     
         14 . The method of  claim 1 , wherein said placental stem cell is a CD10 + , CD34, CD105 + , CD200 +  placental stem cell. 
     
     
         15 . 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. 
     
     
         16 . The method of  claim 14 , wherein said placental stem cells are additionally CD90 +  and CD45 − . 
     
     
         17 . The method of  claim 14 , wherein said placental stem cells are additionally CD80 −  and CD86 − . 
     
     
         18 . A pharmaceutical composition for treating an individual having or at risk of developing a disease, disorder or condition having an inflammatory component, said pharmaceutical composition comprising a therapeutically effective amount of enhanced placental stem cells,
 wherein said cells comprise or have been contacted with an effective amount of modulatory RNA molecules that, when compared to placental stem cells not contacted with said modulatory RNA molecules   (i) suppress an amount of soluble IL-23 protein produced by peripheral blood mononuclear cells (PBMCs) in the presence of said enhanced placental stem cells;   (ii) increase Cox-2 activity in said enhanced placental stem cells;   (iii) increase an amount of PGE2 produced by said enhanced placental stem cells; or   (iv) reduce the level a pro-inflammatory cytokine produced by enhanced placental stein cells,   
       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. 
     
     
         19 . A method of modifying placental stem cells comprising contacting the placental stem cells with an effective amount of modulatory RNA molecules, such that said placental stem cells, after having been contacted with said modulatory RNA molecules
 (i) suppress an amount of soluble IL-23 protein produced by peripheral blood mononuclear cells (PBMCs) in the presence of said placental stem cells;   (ii) increase cyclooxygenase II (Cox-2) activity in said placental stem cells;   (iii) increase an amount of PGE2 produced by said placental stem cells; or   (iv) reduce the level a pro-inflammatory cytokine produced by said placental stem cells,   as compared to an equivalent amount of placental stem cells not contacted with said modulatory RNA molecules.   
     
     
         20 . A composition comprising enhanced placental stem cells, wherein said enhanced placental stem cells have been modified by the method of  claim 19 . 
     
     
         21 . The composition of  claim 20 , w herein said enhanced placental stem cells
 (i) suppress an amount of soluble IL-23 protein produced by peripheral blood mononuclear cells (PBMCs) in the presence of said enhanced placental stem cells;   (ii) increase cyclooxygenase II (Cox-2) activity in said enhanced placental stem cells;   (iii) increase an amount of PGE2 produced by said enhanced placental stem cells; or   (iv) reduce the level a pro-inflammatory cytokine produced by enhanced placental stem cells,   as compared to an equivalent amount of placental stem cells not contacted with said modulatory RNA molecules.   
     
     
         22 . A composition comprising enhanced placental stem cells, wherein said enhanced placental stem cells have been modified by the method of  claim 19 . 
     
     
         23 . A composition comprising enhanced placental stem cells, wherein said enhanced placental stem cells comprises one or more modulatory RNA molecules, and wherein said enhanced placental stem cells
 (i) suppress an amount of soluble IL-23 protein produced by peripheral blood mononuclear cells (PBMCs) in the presence of said enhanced placental stem cells;   (ii) increase cyclooxygenase II (Cox-2) activity in said enhanced placental stem cells;   (iii) increase an amount of PGE2 produced by said enhanced placental stem cells; or   (iv) reduce the level a pro-inflammatory cytokine produced by enhanced placental stem cells,   as compared to an equivalent amount of placental stem cells not contacted with said modulatory RNA molecules.