Histological Localization of Phosphorothioate Oligodeoxynucleotides in Normal Rodent Tissue

Butler, Madeline; Stecker, K.; Bennett, C. Frank

doi:10.1080/07328319708006272

Cited by 84 publications

(125 citation statements)

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“…Hindlimb joints and kidneys were collected on P29, 1 day after the last injection, and processed as described in Materials and Methods. Immunostaining with ASO DNA Ect3 backbone-specific antibody 35 showed the presence of the ASO backbone in the positive control kidney tubules ( Figure 4B) and chondrocytes of the growth plate ( Figures 4D, ASO1, and 4H, ASO2) and articular cartilage ( Figure 4F). No signal was detected in any of these tissues from phosphate buffer (diluent) ( Figures 4A, 4C, 4E, and 4G) or untreated C57BL\6 controls (data not shown).…”

Section: Comp Aso1 Is Delivered To Growth Plate Chondrocytesmentioning

confidence: 99%

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Antisense Reduction of Mutant COMP Reduces Growth Plate Chondrocyte Pathology

et al. 2017

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Mutations in cartilage oligomeric matrix protein cause pseudoachondroplasia, a severe disproportionate short stature disorder. Mutant cartilage oligomeric matrix protein produces massive intracellular retention of cartilage oligomeric matrix protein, stimulating ER and oxidative stresses and inflammation, culminating in post-natal loss of growth plate chondrocytes, which compromises linear bone growth. Treatments for pseudoachondroplasia are limited because cartilage is relatively avascular and considered inaccessible. Here we report successful delivery and treatment using antisense oligonucleotide technology in our transgenic pseudoachondroplasia mouse model. We demonstrate delivery of human cartilage oligomeric matrix protein-specific antisense oligonucleotides to cartilage and reduction of cartilage oligomeric matrix protein expression, which largely alleviates pseudoachondroplasia growth plate chondrocyte pathology. One antisense oligonucleotide reduced steady-state levels of cartilage oligomeric matrix protein mRNA and dampened intracellular retention of mutant cartilage oligomeric matrix protein, leading to a reduction of inflammatory markers and cell death and partial restoration of proliferation. This novel and exciting work demonstrates that antisense-based therapy is a viable approach for treating pseudoachondroplasia and other human cartilage disorders. INTRODUCTIONTreatments of skeletal dysplasia/dwarfing conditions, including pseudoachondroplasia (PSACH), are few and fraught with problems because of the inaccessibility of chondrocytes within the cartilage matrix and the relative avascular nature of the cartilage environment. PSACH is clinically characterized by disproportionate short stature, rhizomelic shortening of the long bones, brachydactyly, and extreme joint laxity. 1,2 Joint pain in childhood and osteoarthritis (OA) in early adulthood are the most debilitating features of PSACH; only symptomatic treatments are available and have limited efficacy. 2,3 The diagnosis is made between 2-3 years of age when linear growth slows and a waddling gait develops. 1,2 Because the diagnosis, in most cases, is made post-natally, treatments aimed at resolving the pathology will have to be started immediately after diagnosis.PSACH is caused by mutations in cartilage oligomeric matrix protein (COMP), a pentameric extracellular matrix (ECM) glycoprotein abundant in musculoskeletal tissues. 4,5 Functionally, COMP facilitates type II collagen fibril assembly, enhances chondrocyte attachment in vitro, and interacts with other ECM proteins, including type II and IX collagens, matrilin 3, and SPARC. [6][7][8][9][10][11][12] In contrast, mutations in COMP cause misfolding of the protein, preventing export from the chondrocyte and resulting in massive retention within the endoplasmic reticulum (ER). 10,[13][14][15] Although this finding has long been appreciated, there was little understanding of the pathologic molecular mechanisms, which are critical to develop mechanism-driven therapeutics. To circumvent this proble...

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Section: Comp Aso1 Is Delivered To Growth Plate Chondrocytesmentioning

confidence: 99%

Section: Comp Aso1 Is Delivered To Growth Plate Chondrocytesmentioning

confidence: 99%

Antisense Reduction of Mutant COMP Reduces Growth Plate Chondrocyte Pathology

et al. 2017

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“…[60][61][62][63] Within the liver, ODNs are not homogenously distributed among different types of liver cells; Kupffer and endothelial cells are predominant sites of receptor mediated ODN internalization. 64,65 However, these cells do not secrete compounds into bile.…”

Section: Discussionmentioning

confidence: 99%

Hepatobiliary transport of bile acid amino acid, bile acid peptide, and bile acid oligonucleotide conjugates in rats

et al. 1999

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Uptake of drugs by bile acid carriers could account for the selectivity of drug actions in the gut and liver. We have previously shown that conjugation of xenobiotics with bile acids facilitates their transfer to hepatocytes and ileal enterocytes. In this study L-alanine and 2 biooligomers, the tetrapeptide L-(ala) 4 and a 15 mer oligodeoxynucleotide (ODN) were coupled covalently via linker molecules to the 3-position of bile acids. The L-alanine-coupled bile acid conjugates were rapidly taken up by the liver and efficiently eliminated into bile. These compounds mimicked hepatic transport of bile acids. Also in case of the tetrapeptide (ala) 4 , bile acid conjugation significantly improved hepatic and intestinal cell uptake and rendered the peptide conjugate resistant to peptidases. Because uptake by isolated hepatocytes was not dependent on sodium ions and was blocked by ochratoxin A, we assume basolateral transport by an oatp-type bile acid carrier. In the case of the 15 mer ODN, normal and bile acid-conjugated oligodeoxynucleotide appeared intact in bile but without marked improvement of hepatocellular uptake and biliary elimination. We conclude that bile acids can deliver small peptides to gut and parenchymal liver cells via bile acid transport pathways, whereas in the case of oligonucleotides an attached bile acid was not sufficient to shuttle them successfully into hepatocytes. (HEPATOLOGY 1999;30:1257-1268

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“…30 Similar to previously published studies, we detected ODNs in the red pulp of the spleen, whereas the white pulp was less efficiently transfected. 31 The red pulp is a highly vascular tissue and allows the passage of blood through the parenchyma, where phagocytic cells remove particulate matter and aged/ abnormal red blood cells. Also in concordance with the literature, we observed ODN uptake in the liver, which is expected given the known role of this organ in ODN clearance.…”

Section: IV Delivery Of Lpdmentioning

confidence: 99%

“…Also in concordance with the literature, we observed ODN uptake in the liver, which is expected given the known role of this organ in ODN clearance. 31,32 Therapeutic relevance ODNs may be therapeutically relevant to CF in three ways. (1) ODNs may be used as decoy molecules to downregulate inflammation.…”

Section: IV Delivery Of Lpdmentioning

confidence: 99%