Wednesday, February 15, 2012

Becoming Taller with MicroRNA

In the future, there are likely to be a number of high tech mechanisms designed to induce height growth.  One of these mechanisms may include microRNA.

MicroRNAs regulate osteogenesis and chondrogenesis.

Being able to induce chondrogenesis is an important ability to induce height growth as you could then form new growth plates.  However, you may still need a demineralized bone surface.

"MicroRNAs (miRNAs) are a class of small molecules and non-coding single strand RNAs that regulate gene expression at the post-transcriptional level by binding to specific sequences within target genes[So we could get miRNAs into the body and then bind to certain genes to induce height growth]. Some miRNAs regulate the proliferation and differentiation of osteoblasts, osteoclasts and chondrocytes, eventually influencing metabolism and bone formation.  Here, we review the recent research progress on the regulation of miRNAs in bone biology, with a particular focus on the miRNA-mediated control mechanisms of bone and cartilage formation."

"[Endochondral ossification] involves the continuous growth and degradation of cartilage and the constant replacement of cartilage tissue by bone tissue."<-Cessation of growth involves the cessation of the growth of cartilage in the bone by getting new cartilage growth whether by LSJL or microRNA, you can get new height growth.

"As members of the Sox family of transcription factors, Sox9, L-Sox5, and Sox6 are necessary for chondrogenic differentiation, and they are expressed through the entire process of chondrogenesis, from mesenchymal condensation to the end of chondrocyte hypertrophy. In all of the cartilage-forming cells in the embryonic stage, Sox9 ensures cell survival and activates Col2a1 and other early cartilage marker genes"<-So we would look for miRNA that upregulates those genes or we would look for exercises/supplements that upregulate those genes.

"miRNA expression has both spatial and temporal specificity as well as tissue or cell specificity."<-thus if you use miRNA's to enhance height growth you have to be mindful of the potential effects on the rest of the body.

"Runx2 and Smads1/5 are necessary for osteogenic differentiation. miR-133 and miR-135 are down-regulated in the BMP-2-induced pluripotent mesenchymal cell line C2C12. Further, miR-133 is a negative regulator of Runx2. miR-133 directly binds to the predicted binding sequence in the 3′ UTR of Runx2 mRNA to inhibit the translation of Runx2 in C2C12 cells. Thus, miR-133 inhibits the osteogenic differentiation of C2C12 that had been induced by BMP-2. Meanwhile, miR-135 represses the osteogenic differentiation of C2C12 cells by targeting the Smad5 gene"<-So, miR-133 and miR-135 are potential targets to repress osteogenic differentiation so stem cells are more likely to differentiate into chondrocytes.

"With microarray technology and luciferase assays, our research group has confirmed that miR-145 acts directly on the 3′ UTR of target gene Sox9. Overexpression of miR-145 decreases the gene expression of MSCs such as Col2a1, Agc1, COMP, Col9a2, and Col11a1, and reduces the contents of GAGs. In contrast, the inhibition of miR-145 could significantly enhance the mRNA expression of the aforementioned genes and promote the contents of GAGs[so we should try to find supplements/exercises that inhibit miR-145]. Adipogenic differentiation marker genes, such as C/EBPδ and C/EBPβ, were not affected by miR-145. It is thus confirmed that Sox9 is the target gene of miR-145 and that miR-145 is a key negative regulator of chondrogenic differentiation, directly targeting Sox9 at an early stage of chondrogenic differentiation. A recent report confirmed the miR-145-mediated regulation of the Sox9 target gene in human articular chondrocytes. The increase in miR-145 could specifically reduce the expression of the cartilage extracellular matrix genes Col2a1 and aggrecan, as well as the tissue-specific miRNAs miR-675 and miR-140."

"cartilage-specific miR-675 could promote cartilage matrix type II collagen expression"<-increasing levels of miR-675 in already existing growth plates may help with height growth.

miR-145 repressions looks the most promising in inducing height growth but it's repression is also associated with several forms of cancer although that may be a symptom rather than the cause of cancer.

Anti-miR-145 may be a potential to lower levels of miR-145.

p53 represses c-Myc through induction of the tumor suppressor miR-145.

"The tumor suppressor p53 negatively regulates a number of genes, including the proto-oncogene c-Myc[Myc may increase height by increasing Lin28 so p53 is bad for height growth]]. One mechanism of the p53-mediated c-Myc repression may involve transcriptional regulation. We show that a putative tumor suppressor, miR-145, is expressed through the phosphoinositide-3 kinase (PI-3K)/Akt and p53 pathways. Importantly, p53 transcriptionally induces the expression of miR-145by interacting with a potential p53 response element (p53RE) in the miR-145 promoter[so inhibiting the p53 pathway may be a way to indirectly affect miR-145]. c-Myc is a direct target for miR-145. Although miR-145 silences the expression of c-Myc, anti-miR-145 enhances its expression. This specific silencing of c-Myc by miR-145 accounts at least in part for the miR-145-mediated inhibition of tumor cell growth both in vitro and in vivo. Finally, the blockade of miR-145 by anti-miR-145 is able to reverse the p53-mediated c-Myc repression. Together, these results define the role of miR-145 in the posttranscriptional regulation of c-Myc by p53 and suggest that, as a new member of the p53 regulatory network, miR-145 provides a direct link between p53 and c-Myc in this gene regulatory network."

pifithrin-α is a potential p53 inhibitor.  So what needs to happen is a supplemental form of Pifithrin-alpha needs to be made and it needs to make sure it doesn't cause cancer.  Likely though, p53 inhibition only accelerates cancer rather than acting as the direct cause.

MicroRNA-337 is associated with chondrogenesis through regulating TGFBR2 expression.

"The expression of six miRNAs in cartilage tissue during development was screened by real-time quantitative polymerase chain reaction (RT-qPCR). Rat models of bone matrix gelatin induced endochondral ossification, collagen-induced arthritis and pristane-induced arthritis were established to examine whether miR-337 is involved in chondrogenesis. Furthermore, the regulation of transforming growth factor-b type II receptor (TGFBR2) expression by miR-337 was determined with the luciferase reporter gene assay and Western blot. The expression of some specific genes relevant to cartilage tissue was tested by RT-qPCR after miR-337 mimic or inhibitor transfection.

MiR-337 expression was significantly down-regulated and almost disappeared in the maturation phases of endochondral ossification. The results of histology and RT-qPCR from three rat models showed that miR-337 is directly bound up with chondrogenesis[thus increasing levels of miR-337 may stimulate chondrogenesis]. Furthermore, the results from the luciferase reporter gene assay and Western blot indicated that miR-337 regulated TGFBR2 expression[Thus TGFBR2 may be key for chondrogenesis as well]. Our study also found that the enhancement of miR-337 may modulate the expression of cartilage-specific genes such as AGC1 in C-28/I2 chondrocytes.

We proved that miRNA-337 is associated with chondrogenesis through regulating TGFBR2 expression, and miRNA-337 can also influence cartilage-specific gene expression in chondrocytes. These findings may provide an important clue for further research in the arthritis pathogenesis and suggest a new remedy for arthritis treatment."

Of course, as always it's possible that miRNA-337 expression is caused by chondrogenesis rather than miRNA-337 causing chondrogenesis.

"AGC1 and ADAMTS4 transcripts in chondrocytes were significantly increased after treatment with miR-337 mimic, while MMP3 mRNA expression was markedly decreased and MMP13 did not obviously change"<-Thus lowering MMP3 levels may help increase height and raising AGC1 and ADAMTS4 levels may help individuals in terms of becoming taller.  According to the NCBI gene back back AGC1 is  a "mitochondrial amino acid transporter, acts both as a glutamate uniporter and as an aspartate-glutamate exchanger; involved in nitrogen metabolism and nitrogen compound biosynthesis".  According to WikiPathways ADAMTS4 is involved in the removal of cartilage matrix.  The full diagram of endochondral ossification on wikipathways can be viewed here.  ADAMTS4 is involved very late in the process of endochondral ossification.  AGC1 is also known as Aggrecan and is also involved late and leads into ADAMTS4 removal of cartilage matrix.

This indicates that removal of the cartilage matrix is very important for height growth which can be evidenced by removal of cartilage matrix being a prerequisite for invasion by osteoblasts.

According to the pathway ADAMTS4 and aggrecan are expressed very late in endochondral ossification however "miR-337 was very low and even hardly detected in the femoral head of adult rats, but highly expressed during the cartilage proliferation".  So miR-337 was evident much earlier.  So perhaps maybe it's MMP-3 expression reduction that was most height increase stimulating.  MMP-3 is not present in the pathway.

"miR-337 activity can promote the ability to anabolism of cartilaginous tissues."<-so miR-337 can help you become taller.

Unfortunately, there is not yet known a way to upregulate miR-337.

The microRNA expression profiles of mouse mesenchymal stem cell during chondrogenic differentiation.

"MiR-140 is tissue-specific expressed in cartilage during embryonic development and play important role in chondrocytes proliferation and differentiaion.It has been validated that histone deacetylase 4 (HDAC4) is its downstream target gene. miR-199 might effect its target gene SMAD1 to regulate chondrogenic differentiation of MSCs"

"miR-140*, miR-140, miR-30a, miR-132/212 and miR143/145 were found that the functions of their potential target genes were correlated with chondrogenic differentiation"

"two up-regulate miRNAs (miR-140*, miR-30a) were associated with decreased expression of their corresponding predicted target mRNA (ADAMTS5, Runx2)."

"two down-regulated miRNAs clusters (miR-132/212, miR-143/145) were associated with increased expression of their corresponding predicted target mRNA (Sox6, ACVR1B)."

Sox9 is associated with miR145.  Sox5 with miR 132/212.

"SMAD1 (miR-30a), SMAD2 (miR-132/212), SMAD3 and SMAD5 (miR-145)"

"suppressed activity of ERK1/2[activated by mir-132/212] resulted in significantly decreased secretion of type II collagen, suggesting ERK1/2 might positively regulate chondrogenesis"

Expression of microRNAs during chondrogenesis of human adipose-derived stem cells.

"Twelve miRNAs were found to be differentially expressed pre- and post- chondrogenic induction by over a two-fold change, including 8 up-regulated miRNAs (miR-193b, miR-199a-3p/hsa-miR-199b-3p, miR-455-3p, miR-210, miR-381, miR-92a, miR-320c, and miR-136), and 4 down-regulated miRNAs (miR-490-5p, miR-4287, miR-BART8*, and miR-US25-1*)"<-different miRNAs than are typically associated with chondrogenic differentiation.

"When mouse mesenchymal stem cells (MSCs) were compared with mature mouse chondrocytes, miR-29a and miR-29b were revealed to directly target 3’ UTR of Col2a1 encoding type II collagen, and their activity was under the regulation of Sox9. miR-199a(*), a bone morphogenic protein 2-responsive microRNA, significantly inhibited early chondrogenesis, as revealed by the reduced expression of early marker genes for chondrogenesis such as cartilage oligomeric matrix protein (COMP), type II collagen, and Sox9, whereas anti-miR-199a(*) increased the expression of these chondrogenic marker genes via direct targeting to smad1 in C3H10T1/2 stem cells"<-so anti-miR-199a could help you grow taller. miR-199a targets Smad1.

"miR-124a was strongly upregulated during chondrogenesis while the expression of miR-96 was substantially suppressed"

"miR-455-3p regulated TGFβ signaling, suppressing the Smad2/3 pathway during chondrognesis"

"[a] predicted candidate target gene, NFKBIA (IKBA), binds NF-κB and inhibits its function"

"the activation of NF-κB was associated with inhibition of chondrogenesis of MSCs by both IL-1β and TNFα in a dose-dependent manner"

"A regulatory circuit comprised of NF-κB, Lin28 {upregulated by LSJL}, Let-7 miRNA, STAT3 and IL-6{upregulated by LSJL}, and a minicircuitry of miR-124, IL6R, STAT3, miR-24, miR-629 and HNF4α {downregulated by LSJL}, was involved in cell transformation"

The role of microRNA-23b in the differentiation of MSC into chondrocyte by targeting protein kinase A signaling.

"both H-89 and microRNA-23b induced differentiation into chondrocyte of hMSCs through down-regulation of protein kinase A (PKA) signaling. The small molecule, H-89, was identified by PCA analysis as a potential mediator of chondrogenic differentiation. H-89 induced the expression of the chondrocyte marker, aggrecan, as well as miR-23b. We searched that miR-23b regulates protein level of PKA. When miR-23b was transfected into hMSCs, chondrogenic differentiation was induced. We confirmed the target of miR-23b using a reporter gene assay. Furthermore, not only H-89 or miR-23b-treated cells, but also cell co-treated with H-89 and miR-23b differentiated into chondrocytes. Our results indicate that H-89 induces the expression of endogenous miR-23b, thereby inducing chondrogenic differentiation by negatively inhibition of PKA signaling."

"miR-145 attenuates differentiation of MSCs into chondrocytes via targeting on the 3′-UTR of Sox9"

LSJL decreases PKIG which inhibits PKA.  However it increases PKIA which also inhibits PKA, so maybe PKA inhibition does play a role in LSJL chondroinduction.

miR-181a promotes osteoblastic differentiation through repression of TGF-β signaling molecules.

"miR-181a [is] highly upregulated during BMP induced osteoblastic differentiation of C2C12 and MC3T3 cells. Overexpression of miR-181a led to upregulation of key markers of osteoblastic differentiation as well as enhanced ALP levels and Alizarin red staining, indicating the importance of this miRNA for osteoblastic differentiation. miR-181 isoforms (181a, 181b, 181c) are expressed during different stages of mouse calvarial and tibial development, implying their role in both endochondral and intramembranous ossification. miR-181a promotes osteoblastic differentiation via repression of TGF-β signaling molecules {So miR-181a is likely anti-chondrogenic therefore miR-181a inhibition is likely pro-chondrogenic} by targeting the negative regulator of osteoblastic differentiation Tgfbi (Tgf-beta induced) and TβR-I/Alk5 (TGF-β type I receptor) {down}. Rgs4 and Gata6 are direct targets of miR-181a."

"miR-181 is upregulated during rat articular cartilage development"

Genes downregulated by miR-181:

More gene expression analysis to be done.

A microRNA signature associated with chondrogenic lineage commitment

"human USSC[unrestricted somatic stem cells] isolation and expansion were carried out"

"downregulation of mir-376a expression and upregulation of mir-630, mir-624 and mir-1268 expression during chondrogenesis. There were 20-fold to 30-fold greater amounts of mir-630 and mir-624 in 21-day chondrocytes compared to undifferentiated USSCs."

"The expression of mir-1268 was 4-fold greater on day 21 than on day 1."

"most of the downregulated miRNAs such as mir-376a, mir-376c, mir-377, mir-337–5p, mir-376a, mir-495, mir-1185, mir-379, mir-127–3p and mir-654–3p are located on chromosome 14. On the other hand, chromosome 19 encodes many of the upregulated miRNAs such as mir-99b, mir-637, mir-181d, mir-515–5p and mir-638."

Anti-let7f increased transcription   AS did pro mir-6230 and mir-624.  Anti mir-221 repressed Sox9.

Mir's good for height growth:

mir-624 genes "SMAD2, WNT5A, PCDH19, CTNNB1"
mir-630 genes "TOB2, GJC1, PDGFRA, TGFBR2"
mir-1268 genes "SOX12, CAMK2G{down in LSJL}"
A more detailed analysis of MiRNA's and target gene comparison to be done.


  1. hey would injectable phytic acid be a useful adjunct for bone matrix demineralization?

  2. well im not an expert but ill try to share informatoon when i can
    it seems really need to find methods to demineralize bone and more information on how to conduct that, i also was thinking perhaps we need to increase the osterocllast activity to demineralize the growth plate for chondrogenesis?

  3. sorry about typo i ment osteoclast,
    the dentin method, is really suprising, perhaps it can be a true alternative to closed plates? should it investigated further? because it seems like best option at the moment to clear the way for stem cells to grow again in plates, get around mineralized area.