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Premature delivery in the domestic sow in response to in utero delivery of AAV9 to fetal piglets.

ABSTRACT: Numerous pediatric neurogenetic diseases may be optimally treated by in utero gene therapy (IUGT); but advancing such treatments requires animal models that recapitulate developmental physiology relevant to humans. One disease that could benefit from IUGT is the autosomal recessive motor neuron disease spinal muscular atrophy (SMA). Current SMA gene-targeting therapeutics are more efficacious when delivered shortly after birth, however postnatal treatment is rarely curative in severely affected patients. IUGT may provide benefit for SMA patients. In previous studies, we developed a large animal porcine model of SMA using AAV9 to deliver a short hairpin RNA (shRNA) directed at porcine survival motor neuron gene (Smn) mRNA on postnatal day 5. Here, we aimed to model developmental features of SMA in fetal piglets and to demonstrate the feasibility of prenatal gene therapy by delivering AAV9-shSmn in utero. Saline (sham), AAV9-GFP, or AAV9-shSmn was injected under direct ultrasound guidance between gestational ages 77-110 days. We developed an ultrasound-guided technique to deliver virus under direct visualization to mimic the clinic setting. Saline injection was tolerated and resulted in viable, healthy piglets. Litter rejection occurred within seven days of AAV9 injection for all other rounds. Our real-world experience of in utero viral delivery followed by AAV9-related fetal rejection suggests that the domestic sow may not be a viable model system for preclinical in utero AAV9 gene therapy studies.

SUBMITTER: Rich KA

PROVIDER: S-EPMC10216882 | biostudies-literature | 2022 Sep

REPOSITORIES: biostudies-literature

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Premature delivery in the domestic sow in response to in utero delivery of AAV9 to fetal piglets.

Rich Kelly A KA Wier Christopher G CG Russo Jessica J Kong Lingling L Heilman Patrick L PL Reynolds Anthony A Knapp Amy A Pino Megan G MG Keckley Elizabeth E Mattox Lori L Malbrue Raphael A RA Sumner Charlotte J CJ Buhimschi Catalin C Kolb Stephen J SJ

Gene therapy 20211122 9

Numerous pediatric neurogenetic diseases may be optimally treated by in utero gene therapy (IUGT); but advancing such treatments requires animal models that recapitulate developmental physiology relevant to humans. One disease that could benefit from IUGT is the autosomal recessive motor neuron disease spinal muscular atrophy (SMA). Current SMA gene-targeting therapeutics are more efficacious when delivered shortly after birth, however postnatal treatment is rarely curative in severely affected ...[more]

PMID: 34803165

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Project description:BackgroundIntrauterine hematopoietic stem cell transplantation (IUT), potentially curative in congenital haematological disease, is often inhibited by deleterious immune responses to donor cells resulting in subtherapeutic donor cell chimerism (DCC). Microchimerism of maternal immune cells (MMc) trafficked into transplanted recipients across the placenta may directly influence donor-specific alloresponsiveness, limiting DCC. We hypothesized that dendritic cells (DC) among trafficked MMc influence the development of tolerogenic or immunogenic responses towards donor cells, and investigated if maternal DC-depletion reduced recipient alloresponsiveness and enhanced DCC.MethodsUsing transgenic CD11c.DTR (C57BL/6) female mice enabled transient maternal DC-depletion with a single dose of diphtheria toxin (DT). CD11c.DTR females and BALB/c males were cross-mated, producing hybrid pups. IUT was performed at E14 following maternal DT administration 24 h prior. Bone marrow-derived mononuclear cells were transplanted, obtained from semi-allogenic BALB/c (paternal-derived; pIUT), C57BL/6 (maternal-derived; mIUT), or fully allogenic (aIUT) C3H donor mice. Recipient F1 pups were analyzed for DCC, while maternal and IUT-recipient immune cell profile and reactivity were examined via mixed lymphocyte reactivity functional assays. T- and B-cell receptor repertoire diversity in maternal and recipient cells were examined following donor cell exposure.ResultsDCC was highest and MMc was lowest following pIUT. In contrast, aIUT recipients had the lowest DCC and the highest MMc. In groups that were not DC-depleted, maternal cells trafficked post-IUT displayed reduced TCR & BCR clonotype diversity, while clonotype diversity was restored when dams were DC-depleted. Additionally, recipients displayed increased expression of regulatory T-cells and immune-inhibitory proteins, with reduced proinflammatory cytokine and donor-specific antibody production. DC-depletion did not impact initial donor chimerism. Postnatal transplantation without immunosuppression of paternal donor cells did not increase DCC in pIUT recipients; however there were no donor-specific antibody production or immune cell changes.ConclusionsThough maternal DC depletion did not improve DCC, we show for the first time that MMc influences donor-specific alloresponsiveness, possibly by expanding alloreactive clonotypes, and depleting maternal DC promotes and maintains acquired tolerance to donor cells independent of DCC, presenting a novel approach to enhancing donor cell tolerance following IUT. This may have value when planning repeat HSC transplantations to treat haemoglobinopathies.

Project description:Maternal care behaviour is crucial for offspring quality and survival in pigs. Defining care is therefore essential for ensuring the welfare of pigs and sustainability of pig production. The aim of the present study was to investigate the relationship between sow nest building, communication with piglets (sniffing, nudging, grunting) during resting and activity, and piglet survival in three different sow breeds: a maternal line selected for high weaned pig production (Landrace), a paternal line selected for meat traits (Duroc), and a crossbred line (Landrace and Yorkshire). We predicted that a higher frequency of nest building and sow communication would have a positive impact on piglet survival. Secondly, we predicted that a high level of maternal care outside the time of nursing (nest building and communication) would increase the quality of the litter (weight at weaning). We also predicted that nest building activity and sow communication would be more pronounced in maternal sow lines selected for maternal traits than in a non-selected, paternal line, and that primiparous sows would perform more nest building behaviour and communicate more than multiparous sows due to high investment in their first litter. Finally, an impaired condition around farrowing (i.e. low body condition score and presence of shoulder lesions) was predicted to be negatively correlated to care behaviours. Data was collected on 38 sows with 511 born piglets. Sows with their litters, were loose-housed in individual farrowing pens until weaning. Nest building activity can be partly considered as maternal care behaviour as it prepares the sows for motherhood and is associated with a lower proportion of stillborn piglets (P < 0.001), starved piglets (P = 0.004), and overlaid piglets (P = 0.034). As predicted, sows that communicated more while being active had lower postnatal piglet mortality (starvation (P < 0.001), less overlying (P < 0.001), overlying with (P < 0.001), and without the milk in the stomach (P < 0.001) and fewer that died of other causes (P < 0.001), higher piglet survival (P < 0.001) and litter weight (P < 0.001) at weaning irrespective of the breed. A higher level of communication while active was associated with more pronounced shoulder lesions in sows (P = 0.010), suggesting a positive association between good maternal care and prevalence of shoulder lesions. We also found that resting sows that communicated more with piglets outside the time of nursing, had higher postnatal piglet mortality (P < 0.001) due to starvation (P < 0.001), overlying (P < 0.001), overlying with (P < 0.001), or without milk (P < 0.001). Communication during resting was more pronounced with increasing litter size at birth (P < 0.001), especially for thin sows (P < 0.001). Communication during resting was more pronounced in the non-selected Duroc line (P < 0.001). Our results suggest that sow communication while being active is a good predictor of good maternal care, piglet survival and litter quality in three different breeds of domestic pigs.

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Premature delivery in the domestic sow in response to in utero delivery of AAV9 to fetal piglets.

Publications

Premature delivery in the domestic sow in response to in utero delivery of AAV9 to fetal piglets.

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