Gene Delivery to Intestinal Epithelial Cells In vitro and In vivo with Recombinant Adeno-Associated Virus Types 1, 2 and 5

Abstract

Intestinal disorders such as inflammatory bowel disease (IBD) result in chronic illness requiring lifelong therapy. Our aim was to evaluate the efficacy of recombinant adeno-associated virus (AAV) vector-mediated gene delivery to intestinal epithelial cells in vitro and in vivo. Human colon epithelial cell lines and colon biopsies were transduced using AAV pseudotypes 2/1, 2/2, and 2/5 encoding green fluorescence protein (GFP). Mice were administered the same vectors through oral, enema, intraperitoneal (IP) injection and superior mesenteric artery (SMA) injection routes. Tropism and efficiency were determined by microscopy, flow cytometry, immunohistochemistry and PCR. Caco2 cells were more permissive to AAV transduction. Human colon epithelial cells in organ culture were more effectively transduced by AAV2/2. SMA injection provided the most effective means of vector gene transfer to small intestine and colonic epithelial cells in vivo. NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author Manuscript applicable for the investigation of IBD pathogenesis and novel therapeutic options, but also revealed the need for further studies to identify more efficient pseudotypes.

Human Galectin 3 Binding Protein Interacts with Recombinant Adeno-Associated Virus Type 6

“…Furthermore, interaction of hu-G3BP with rAAV-6 led to the formation of aggregates and hampered transduction when the two were codelivered into the mouse. Based on these data, we propose that species-specific interactions of AAVs with blood proteins may differentially impact vector distribution and efficacy in different animal models.T he recombinant adeno-associated vector (rAAV) platform, derived from a nonpathogenic dependovirus, has many attributes suitable for in vivo gene transfer: rAAV vectors are capable of transducing a wide range of cell types, including dividing and nondividing cells; rAAV genomes persist as episomal chromatin in the nucleus of transduced cells (38); and stable, persistent expression has been reported for many transgenes in different tissues and species (6,12,36,39). rAAVs have proven to be efficient in preclinical studies in animal models (16,28), and results from clinical trials are promising (7,47).…”

Targeting murine small bowel and colon through selective superior mesenteric artery injection

“…Previous attempts at delivery of agents to the intestinal epithelium via oral, intraperitoneal injection, and systemic injection have not yielded good results. 3,6,11,16 Administration of insoluble agents by rectal enema have demonstrated uptake in the rectum only and thus is not a optimal approach for broader colonic administration. 17 Although the isolated SMA injection technique effectively reached the small bowel and colon, these data also demonstrate that there is systemic escape, albeit in low levels.…”

“…Sferra et al 8 reported the successful SMA injection in a rat for vector administration and thus we developed and reported the first isolated SMA injection in a mouse model. 11 To improve our efficiency and hasten postoperative recovery we have developed a modified microsurgical approach. Here we report the detailed steps for this surgical approach to verify the anatomical distribution, evaluate the biodistribution, and examine the feasibility of administering biological agents following SMA injection.…”

In vivo tissue-tropism of adeno-associated viral vectors

“…Successful transduction of the rabbit lung [61,62], and the rat carotid arteries [63] by AAV2 vectors was also reported in 1997. Subsequently, various AAV serotype vector-mediated gene transfer in various cell cells and tissues, such as intestinal epithelial cells [64], pancreatic beta cells [65], salivary glands [66], maxillary sinus [67] and temporomandibular joints [68], and in various animal models, such as hamster [69], Japanese quail [70], gerbil [71], cat [72], dog [73], and cynomolgus monkeys [74] and were also reported. A representative example of some of these studies with the various AAV serotype vectors in the most commonly used animal models is depicted in Figure 1.…”