Severe Developmental B Lymphopoietic Defects in Foxp3-Deficient Mice are Refractory to Adoptive Regulatory T Cell Therapy

The role of Foxp3-expressing regulatory T (Treg) cells in tolerance and autoimmunity is well-established. However, although of considerable clinical interest, the role of Treg cells in the regulation of hematopoietic homeostasis remains poorly understood. Thus, we analysed B and T lymphopoiesis in the scurfy (Sf) mouse model of Treg cell deficiency. In these experiments, the near-complete block of B lymphopoiesis in the BM of adolescent Sf mice was attributed to autoimmune T cells. We could exclude a constitutive lympho-hematopoietic defect or a B cell-intrinsic function of Foxp3. Efficient B cell development in the BM early in ontogeny and pronounced extramedullary B lymphopoietic activity resulted in a peripheral pool of mature B cells in adolescent Sf mice. However, marginal zone B and B-1a cells were absent throughout ontogeny. Developmental B lymphopoietic defects largely correlated with defective thymopoiesis. Importantly, neonatal adoptive Treg cell therapy suppressed exacerbated production of inflammatory cytokines and restored thymopoiesis but was ineffective in recovering defective B lymphopoiesis, probably due to a failure to compensate production of stroma cell-derived IL-7 and CXCL12. Our observations on autoimmune-mediated incapacitation of the BM environment in Foxp3-deficient mice will have direct implications for the rational design of BM transplantation protocols for patients with severe genetic deficiencies in functional Foxp3+ Treg cells

Integrated Solutions for the Water-Energy-Land Nexus: Are Global Models Rising to the Challenge?

Increasing human demands for water, energy, food and materials, are expected to accentuate resource supply challenges over the coming decades. Experience suggests that long-term strategies for a single sector could yield both trade-offs and synergies for other sectors. Thus, long-term transition pathways for linked resource systems should be informed using nexus approaches. Global integrated assessment models can represent the synergies and trade-offs inherent in the exploitation of water, energy and land (WEL) resources, including the impacts of international trade and climate policies. In this study, we review the current state-of-the-science in global integrated assessment modeling with an emphasis on how models have incorporated integrated WEL solutions. A large-scale assessment of the relevant literature was performed using online databases and structured keyword search queries. The results point to the following main opportunities for future research and model development: (1) improving the temporal and spatial resolution of economic models for the energy and water sectors; (2) balancing energy and land requirements across sectors; (3) integrated representation of the role of distribution infrastructure in alleviating resource challenges; (4) modeling of solution impacts on downstream environmental quality; (5) improved representation of the implementation challenges stemming from regional financial and institutional capacity; (6) enabling dynamic multi-sectoral vulnerability and adaptation needs assessment; and (7) the development of fully-coupled assessment frameworks based on consistent, scalable, and regionally-transferable platforms. Improved database management and computational power are needed to address many of these modeling challenges at a global-scale

CUX1-related neurodevelopmental disorder: deep insights into phenotype-genotype spectrum and underlying pathology

Heterozygous, pathogenic CUX1 variants are associated with global developmental delay or intellectual disability. This study delineates the clinical presentation in an extended cohort and investigates the molecular mechanism underlying the disorder in a Cux1+/− mouse model. Through international collaboration, we assembled the phenotypic and molecular information for 34 individuals (23 unpublished individuals). We analyze brain CUX1 expression and susceptibility to epilepsy in Cux1+/− mice. We describe 34 individuals, from which 30 were unrelated, with 26 different null and four missense variants. The leading symptoms were mild to moderate delayed speech and motor development and borderline to moderate intellectual disability. Additional symptoms were muscular hypotonia, seizures, joint laxity, and abnormalities of the forehead. In Cux1+/− mice, we found delayed growth, histologically normal brains, and increased susceptibility to seizures. In Cux1+/− brains, the expression of Cux1 transcripts was half of WT animals. Expression of CUX1 proteins was reduced, although in early postnatal animals significantly more than in adults. In summary, disease-causing CUX1 variants result in a non-syndromic phenotype of developmental delay and intellectual disability. In some individuals, this phenotype ameliorates with age, resulting in a clinical catch-up and normal IQ in adulthood. The post-transcriptional balance of CUX1 expression in the heterozygous brain at late developmental stages appears important for this favorable clinical course.CAG was supported by the Eunice Kennedy Shriver National Institute Of Child Health & Human Development of the National Institutes of Health under Award Number P50 HD103525. This work was funded by PID2020-112831GB-I00 AEI /10.13039/501100011033 (MN). SS was supported by a grant from the NIH/NINDS (K23NS119666). SWS is supported by the Hospital for Sick Children Foundation, Autism Speaks, and the University of Toronto McLaughlin Center. EM-G was supported by a grant from MICIU FPU18/06240. EVS. was supported by a grant from the NIH (EY025718). CRF was supported by the fund to support clinical research careers in the Region of Southern Denmark (Region Syddanmarks pulje for kliniske forskerkarriereforløb).Peer reviewe

Integrated Solutions for the Water-Energy-Land Nexus: Are Global Models Rising to the Challenge?

Increasing human demands for water, energy, food and materials, are expected to accentuate resource supply challenges over the coming decades. Experience suggests that long-term strategies for a single sector could yield both trade-offs and synergies for other sectors. Thus, long-term transition pathways for linked resource systems should be informed using nexus approaches. Global integrated assessment models can represent the synergies and trade-offs inherent in the exploitation of water, energy and land (WEL) resources, including the impacts of international trade and climate policies. In this study, we review the current state-of-the-science in global integrated assessment modeling with an emphasis on how models have incorporated integrated WEL solutions. A large-scale assessment of the relevant literature was performed using online databases and structured keyword search queries. The results point to the following main opportunities for future research and model development: (1) improving the temporal and spatial resolution of economic models for the energy and water sectors; (2) balancing energy and land requirements across sectors; (3) integrated representation of the role of distribution infrastructure in alleviating resource challenges; (4) modeling of solution impacts on downstream environmental quality; (5) improved representation of the implementation challenges stemming from regional financial and institutional capacity; (6) enabling dynamic multi-sectoral vulnerability and adaptation needs assessment; and (7) the development of fully-coupled assessment frameworks based on consistent, scalable, and regionally-transferable platforms. Improved database management and computational power are needed to address many of these modeling challenges at a global-scale

RA treatment leads to up-regulation of gut homing molecules on the surface of cultured PEC B cells.

<p>Expression of surface α4β7 and CCR9 on IgA^- PEC (A) or splenic (B) B cells sorted in fashion similar to previous experiments was checked by flow cytometry after four days of culture under various combinations of stimulatory conditions. Using the same cultures (as in A and B), expression of surface IgA and IgM on PEC (C) and splenic (D) B cells was checked by flow cytometry. Data show the results of one of two independent experiments.</p

TGF-β enhances IgA switching among PEC B1b cells <i>in vitro</i>.

<p>Results of IgA and IgM ELISPOT assays performed in triplicates on sort purified PEC B cell subpopulations after 4 days of treatment with the indicated combination of stimulatory factors in culture. Surface IgA⁺ cells were excluded by sorting from B1a (IgA^-CD19^hiCD5⁺CD43⁺Mac-1⁺), B1b (IgA^-CD19^hiCD5^-CD43⁺Mac-1⁺) and B2 (IgA^-CD19^loCD5^-CD43^-Mac-1^-) B cell subpopulations. Data show the results of one of three independent experiments. Bars represent mean ± SD.</p

RA and TGF-β treated PEC B1 cells migrate to spleen and gut and produce serum and intestinal secretory IgA.

<p>IgA^- PEC B1a and B1b cells sorted in a fashion similar to previous experiments were cultured under various IgA CSR inducing conditions for 3 days and transferred intraperitoneally (16000 live cells/mouse) into lymphopenic Rag1^-/- recipients. (A) Unstimulated splenocytes and 2 days LPS (25 µg/ml) stimulated PECs of the recipient mice were analyzed for the presence of IgA and IgM producing cells by ELISPOT 2 weeks after adoptive B cell transfer. (B) Levels of secretory Igs in the serum and gut lavage of recipient mice were determined by ELISA. Per group, 2-3 mice were used as recipients. Cells pooled from the recipients belonging to the same group were used for ELISPOT assay and it was done in triplicates. Data show the results of one of two independent experiments. Bars represent mean ± SD.</p

PEC and splenic B1 cell derived IgA VH sequences display high frequencies of nucleotide exchanges due to somatic hypermutation.

<p>Frequency of mutations observed in the IgM or IgA VH region sequences derived from PEC and splenic B1 cells cultured for 4 days with the indicated combinations of IgA CSR inducing factors. (A) Each colored sector represents a particular number of mutations found at a particular frequency. Total number of sequences is shown in the middle. (B) Number of replacement and silent mutations amongst IgM and IgA VH sequences derived from indicated B cell populations. Sequences were analyzed using SEQUENCHER™ Version 4.1 for Macintosh software (Gene Codes Corporation). Assignment of VH chain was done using VBASE2 database (<a href="http://www.vbase2.org/" target="_blank">http://www.vbase2.org/</a>). Only functional sequences were included for the mutation analysis. Bars represent mean ± SD.</p

TGF-β in combination with RA synergistically induces IgA switching by PEC B cells.

<p>(A) Results of IgA and IgM ELISPOT assays performed in triplicates on sort purified PEC B cell subpopulations after 4 days of treatment with the indicated combination of stimulatory factors in culture. Surface IgA⁺ cells were excluded by sorting from B1a (IgA^-CD19^hiCD5⁺CD43⁺Mac-1⁺), B1b (IgA^-CD19^hiCD5^-CD43⁺Mac-1⁺) and B2 (IgA^-CD19^loCD5^-CD43^-Mac-1^-) B cell subpopulations. (B) Amount of secretory Igs determined by ELISA in the supernatant of B cells cultured for 4 days under the indicated stimulatory conditions. Data show the results of one of two independent experiments. Bars represent mean ± SD.</p

Increase in cell numbers after 4 days of culture under various conditions.

<p>¹ Sorted IgA^- PEC and splenic B cells were cultured under different conditions for 4 days and the fold increase in cell number was calculated by dividing the cell count after 4 days by the number of cells at the beginning of the cell culture. The values represent mean ± SD calculated from three independent culture experiments.</p