Nanocarrier lipid composition modulates the impact of pulmonary surfactant protein B (SP-B) on cellular delivery of siRNA

Two decades since the discovery of the RNA interference (RNAi) pathway, we are now witnessing the approval of the first RNAi-based treatments with small interfering RNA (siRNA) drugs. Nevertheless, the widespread use of siRNA is limited by various extra- and intracellular barriers, requiring its encapsulation in a suitable (nanosized) delivery system. On the intracellular level, the endosomal membrane is a major barrier following endocytosis of siRNA-loaded nanoparticles in target cells and innovative materials to promote cytosolic siRNA delivery are highly sought after. We previously identified the endogenous lung surfactant protein B (SP-B) as siRNA delivery enhancer when reconstituted in (proteo) lipid-coated nanogels. It is known that the surface-active function of SP-B in the lung is influenced by the lipid composition of the lung surfactant. Here, we investigated the role of the lipid component on the siRNA delivery-promoting activity of SP-B proteolipid-coated nanogels in more detail. Our results clearly indicate that SP-B prefers fluid membranes with cholesterol not exceeding physiological levels. In addition, SP-B retains its activity in the presence of different classes of anionic lipids. In contrast, comparable fractions of SP-B did not promote the siRNA delivery potential of DOTAP:DOPE cationic liposomes. Finally, we demonstrate that the beneficial effect of lung surfactant on siRNA delivery is not limited to lung-related cell types, providing broader therapeutic opportunities in other tissues as well

Nanocarrier lipid composition modulates the impact of pulmonary surfactant protein B (SP-B) on cellular delivery of siRNA

Two decades since the discovery of the RNA interference (RNAi) pathway, we are now witnessing the approval of the first RNAi-based treatments with small interfering RNA (siRNA) drugs. Nevertheless, the widespread use of siRNA is limited by various extra- and intracellular barriers, requiring its encapsulation in a suitable (nanosized) delivery system. On the intracellular level, the endosomal membrane is a major barrier following endocytosis of siRNA-loaded nanoparticles in target cells and innovative materials to promote cytosolic siRNA delivery are highly sought after. We previously identified the endogenous lung surfactant protein B (SP-B) as siRNA delivery enhancer when reconstituted in (proteo) lipid-coated nanogels. It is known that the surface-active function of SP-B in the lung is influenced by the lipid composition of the lung surfactant. Here, we investigated the role of the lipid component on the siRNA delivery-promoting activity of SP-B proteolipid-coated nanogels in more detail. Our results clearly indicate that SP-B prefers fluid membranes with cholesterol not exceeding physiological levels. In addition, SP-B retains its activity in the presence of different classes of anionic lipids. In contrast, comparable fractions of SP-B did not promote the siRNA delivery potential of DOTAP:DOPE cationic liposomes. Finally, we demonstrate that the beneficial effect of lung surfactant on siRNA delivery is not limited to lung-related cell types, providing broader therapeutic opportunities in other tissues as well

Architectural Root Responses of Rice to Reduced Water Availability Can Overcome Phosphorus Stress

Drought and low phosphorus (P) availability are major limitations for rainfed rice production. Crop roots are important for soil resource acquisition and tolerance to P and water limitations. Two pot and two field trials were conducted to evaluate architectural root responses of contrasting rice varieties to combinations of different levels of P (deficient to non-limiting) and water availability (water stressed to submergence) and to identify the interactions with different varieties. Root development was then related to drought and/or low P tolerance. Although shoot and root growth responded more to P than to water availability, architectural root responses to water were much more prominent than responses to P availability. Reduced water availability decreased nodal thickness and increased secondary root branching, both factors partially enhancing P uptake efficiency and even overcoming a decreased root:shoot ratio under reduced water availability. In contrast to root thickness and secondary branching, basal lateral root density was strongly determined by variety and was related to enhanced P uptake. Reduced water availability induces root modifications which—apart from enhancing drought resilience—also affect P uptake efficiency. Future research on rice roots and nutrient uptake may hence take into account the large effects of water on root development

Architectural Root Responses of Rice to Reduced Water Availability Can Overcome Phosphorus Stress

© 2018 by the authors. Drought and low phosphorus (P) availability are major limitations for rainfed rice production. Crop roots are important for soil resource acquisition and tolerance to P and water limitations. Two pot and two field trials were conducted to evaluate architectural root responses of contrasting rice varieties to combinations of different levels of P (deficient to non-limiting) and water availability (water stressed to submergence) and to identify the interactions with different varieties. Root development was then related to drought and/or low P tolerance. Although shoot and root growth responded more to P than to water availability, architectural root responses to water were much more prominent than responses to P availability. Reduced water availability decreased nodal thickness and increased secondary root branching, both factors partially enhancing P uptake efficiency and even overcoming a decreased root:shoot ratio under reduced water availability. In contrast to root thickness and secondary branching, basal lateral root density was strongly determined by variety and was related to enhanced P uptake. Reduced water availability induces root modifications which—apart from enhancing drought resilience—also affect P uptake efficiency. Future research on rice roots and nutrient uptake may hence take into account the large effects of water on root development.status: publishe

Water and phosphorus uptake by upland rice root systems unraveled under multiple scenarios: linking a 3D soil-root model and data

Background and aims Upland rice is often grown where water and phosphorus (P) are limited and these two factors interact on P bioavailability. To better understand this interaction, mechanistic models representing small-scale nutrient gradients and water dynamics in the rhizosphere of full-grown root systems are needed.Methods Rice was grown in large columns using a P-deficient soil at three different P supplies in the topsoil (deficient, suboptimal, non-limiting) in combination with two water regimes (field capacity versus drying periods). Root architectural parameters and P uptake were determined. Using a multiscale model of water and nutrient uptake, in-silico experiments were conducted by mimicking similar P and water treatments. First, 3D root systems were reconstructed by calibrating an architecure model with observed phenological root data, such as nodal root number, lateral types, interbranch distance, root diameters, and root biomass allocation along depth. Secondly, the multiscale model was informed with these 3D root architectures and the actual transpiration rates. Finally, water and P uptake were simulated.Key results The plant P uptake increased over threefold by increasing P and water supply, and drying periods reduced P uptake at high but not at low P supply. Root architecture was significantly affected by the treatments. Without calibration, simulation results adequately predicted P uptake, including the different effects of drying periods on P uptake at different P levels. However, P uptake was underestimated under P deficiency, a process likely related to an underestimated affinity of P uptake transporters in the roots. Both types of laterals (i.e. S- and L-type) are shown to be highly important for both water and P uptake, and the relative contribution of each type depend on both soil P availability and water dynamics. Key drivers in P uptake are growing root tips and the distribution of laterals.Conclusions This model-data integration demonstrates how multiple co-occurring single root phene responses to environmental stressors contribute to the development of a more efficient root system. Further model improvements such as the use of Michaelis constants from buffered systems and the inclusion of mycorrhizal infections and exudates are proposed

A functional–structural model of upland rice root systems reveals the importance of laterals and growing root tips for phosphate uptake from wet and dry soils

Background and AimsUpland rice is often grown where water and phosphorus (P) are limited. To better understand the interaction between water and P availability, functional–structural models that mechanistically represent small-scale nutrient gradients and water dynamics in the rhizosphere are needed.MethodsRice was grown in large columns using a P-deficient soil at three P supplies in the topsoil (deficient, sub-optimal and non-limiting) in combination with two water regimes (field capacity vs. drying periods). Root system characteristics, such as nodal root number, lateral types, interbranch distance, root diameters and the distribution of biomass with depth, as well as water and P uptake, were measured. Based on the observed root data, 3-D root systems were reconstructed by calibrating the structural architecure model CRootBox for each scenario. Water flow and P transport in the soil to each of the individual root segments of the generated 3-D root architectures were simulated using a multiscale flow and transport model. Total water and P uptake were then computed by adding up the uptake by all the root segments.Key ResultsMeasurements showed that root architecture was significantly affected by the treatments. The moist, high P scenario had 2.8 times the root mass, double the number of nodal roots and more S-type laterals than the dry, low P scenario. Likewise, measured plant P uptake increased >3-fold by increasing P and water supply. However, drying periods reduced P uptake at high but not at low P supply. Simulation results adequately predicted P uptake in all scenarios when the Michaelis–Menten constant (Km) was corrected for diffusion limitation. They showed that the key drivers for P uptake are the different types of laterals (i.e. S- and L-type) and growing root tips. The L-type laterals become more important for overall water and P uptake than the S-type laterals in the dry scenarios. This is true across all the P treatments, but the effect is more pronounced as the P availability decreases.ConclusionsThis functional–structural model can predict the function of specific rice roots in terms of P and water uptake under different P and water supplies, when the structure of the root system is known. A future challenge is to predict how the structure root systems responds to nutrient and water availability

A functional–structural model of upland rice root systems reveals the importance of laterals and growing root tips for phosphate uptake from wet and dry soils

status: Published onlin

Anatomical root responses of rice to combined phosphorus and water stress – relations to tolerance and breeding opportunities

status: Published onlin