Papers by Allan Goldstein
Biomolecules
With recent technical advances and diminishing sequencing costs, single-cell sequencing modalitie... more With recent technical advances and diminishing sequencing costs, single-cell sequencing modalities have become commonplace. These tools permit analysis of RNA expression, DNA sequence, chromatin structure, and cell surface antigens at single-cell resolution. Simultaneous measurement of numerous parameters can resolve populations including rare cells, thus revealing cellular diversity within organs and permitting lineage reconstruction in developing tissues. Application of these methods to the enteric nervous system has yielded a wealth of data and biological insights. We review recent papers applying single-cell sequencing tools to the nascent neural crest and to the developing and mature enteric nervous system. These studies have shown significant diversity of enteric neurons and glia, suggested paradigms for neuronal specification, and revealed signaling pathways active during development. As technology evolves and multiome techniques combining two or more of transcriptomic, genom...
Clinics in colon and rectal surgery, 2018
Constipation is a common childhood problem, but an anatomic or physiologic cause is identified in... more Constipation is a common childhood problem, but an anatomic or physiologic cause is identified in fewer than 5% of children. By definition, idiopathic constipation is a diagnosis of exclusion. Careful clinical evaluation and thoughtful use of imaging and other testing can help exclude specific causes of constipation and guide therapy. Medical management with laxatives is effective for the majority of constipated children. For those patients unresponsive to medications, however, several surgical options can be employed, including anal procedures, antegrade colonic enemas, colorectal resection, and intestinal diversion. Judicious use of these procedures in properly selected patients and based on appropriate preoperative testing can lead to excellent outcomes. This review summarizes the surgical options available for managing refractory constipation in children and provides guidance on how to choose the best procedure for a given patient.
Nature communications, Oct 10, 2017
Patients with short bowel syndrome lack sufficient functional intestine to sustain themselves wit... more Patients with short bowel syndrome lack sufficient functional intestine to sustain themselves with enteral intake alone. Transplantable vascularized bioengineered intestine could restore nutrient absorption. Here we report the engineering of humanized intestinal grafts by repopulating decellularized rat intestinal matrix with human induced pluripotent stem cell-derived intestinal epithelium and human endothelium. After 28 days of in vitro culture, hiPSC-derived progenitor cells differentiate into a monolayer of polarized intestinal epithelium. Human endothelial cells seeded via native vasculature restore perfusability. Ex vivo isolated perfusion testing confirms transfer of glucose and medium-chain fatty acids from lumen to venous effluent. Four weeks after transplantation to RNU rats, grafts show survival and maturation of regenerated epithelium. Systemic venous sampling and positron emission tomography confirm uptake of glucose and fatty acids in vivo. Bioengineering intestine on ...
The Mechanism of Excessive Intestinal Inflammation in Necrotizing Enterocolitis: An Immature Innate Immune Response
PLoS ONE, 2011
Macromolecular bioscience, Jan 2, 2015
Mucus forms a protective hydrogel layer over the intestinal epithelium, presenting a selective an... more Mucus forms a protective hydrogel layer over the intestinal epithelium, presenting a selective and robust barrier to the uptake of particulates and microbe invasion. Disease can alter mucus production and composition, thus potentially modifying mucosal barrier properties. Hirschsprung's disease (HD) is a developmental abnormality of the nervous system often complicated by intestinal infection. An investigation of colonic mucus barrier properties in an HD animal model, endothelin receptor B mutant mice, revealed significantly reduced microsphere (passive) and microbe (active) transport rates (7-fold and 3.6-fold, respectively, in proximal colonic mucus) relative to wild-type. Transport differences were evident in both the ganglionic and aganglionic colon segments, in agreement with the risk of HD-associated enterocolitis after surgery to remove aganglionic colon segments. The development of therapies aimed at altering colonic mucus barrier properties could be explored towards pre...
Textbook of Clinical Pediatrics, 2012
Table 418.1 Causes of neonatal intestinal obstruction Mechanical causes Functional causes Malrota... more Table 418.1 Causes of neonatal intestinal obstruction Mechanical causes Functional causes Malrotation with midgut volvulus Prematurity Duodenal atresia Sepsis Annular pancreas Maternal narcotic use Jejunoileal atresia Magnesium use Colonic atresia Congenital hypothyroidism Meconium ileus Small left colon syndrome Anorectal malformation Meconium plug syndrome Presacral mass Hirschsprung's disease 4012 418 Congenital Intestinal Obstruction
New England Journal of Medicine, 2012
The patient was born at another hospital to a teenaged primigravida by vaginal delivery after a f... more The patient was born at another hospital to a teenaged primigravida by vaginal delivery after a full-term, uncomplicated gestation. The mother had received prenatal care; she had no history of sexually transmitted infections, and prenatal screening tests were negative. Meconium was present at delivery. The patient's birth weight was 4.3 kg (95th percentile), and the 1-minute and 5-minute Apgar scores were 7 and 9, respectively. Breast-feeding was initiated. The newborn passed two stools on the second day (the first at 30 hours of age). The next day, he reportedly had one loose, green stool. He was discharged home at 50 hours of age. He lived with his mother and her parents. When he was 3 and 4 days of age, he vomited yellow-green emesis on several occasions. He was otherwise well and breastfeeding regularly. At 3 a.m. on the day of admission, he became fussy and did not complete his normal feeding. Between 3 a.m. and 9 a.m., approximately six episodes of vomiting (initially breast milk, followed by mucus) occurred, with increasingly foul-smelling emesis. Diarrhea developed, and urine output decreased. Later that morning, he became less active; abdominal distention developed, and he stopped voiding. He was taken to a clinic affiliated with this hospital. On examination, he appeared tired, with intermittent grunting. The rectal temperature was 38.0°C. The abdomen was distended and tender, with hypoactive bowel sounds; stool was positive for occult blood. He was transported by ambulance to the emergency department of this hospital. On examination, the patient appeared alert and slightly uncomfortable. The temperature was 37.2°C, the pulse 160 beats per minute, the respiratory rate 30 breaths per minute, and the oxygen saturation 100% while he was breathing ambient air. The weight was 3.9 kg. The abdomen was distended, soft, and tympanic, with decreased bowel sounds. There were small, bilateral noncommunicating hydroceles. The remainder of the examination was normal. A stool specimen showed no occult blood. Urinalysis revealed clear orange urine, with a specific gravity greater than 1.030, pH 6.5, nitrites, 1+ urobilinogen, 3+ bilirubin, 2+ albumin, and trace white cells, blood, glucose, and ketones. Urinalysis also revealed 0 to 2 red cells, 3 to 5 white
Neurology, 1999
To determine whether individuals with situs inversus totalis (SI), a condition in which there is ... more To determine whether individuals with situs inversus totalis (SI), a condition in which there is a mirror-image reversal of asymmetric visceral organs, have alterations in brain asymmetries. Background: The human brain is asymmetric in structure and function. Although correlations between anatomic asymmetries and functional lateralization in human brain have been demonstrated, it has been difficult to further analyze them. Characterization of asymmetries of brain structure and function in SI might advance the understanding of these relationships. Methods: Using anatomic and functional MRI techniques, we analyzed asymmetries in the brains of three individuals with SI. Results: Two major anatomic asymmetries of the cerebral hemispheres, the frontal and occipital petalia, were reversed in individuals with SI. In contrast, SI subjects had left cerebral hemisphere language dominance on functional MRI analysis as well as strong right-handedness. Conclusion: These observations suggest that the developmental factors determining anatomic asymmetry of the cerebral petalia and viscera are distinct from those producing the functional lateralization of language.
Neurogastroenterology & Motility, 2013
Background-The metabolic pathways associated with colonic motility are unknown. To identify poten... more Background-The metabolic pathways associated with colonic motility are unknown. To identify potential metabolic targets for treatment of constipation, we examined the metabolic profile before and after a meal challenge in a cohort of children with constipation and determined its relationship with postprandial colon motility patterns. Methods-In this prospective study, 187 metabolites were measured by liquid chromatographymass spectrometry at multiple time points before and after a standardized meal in constipated children undergoing a colon manometry. Postprandial metabolite levels were compared with baseline and also correlated with multiple manometric measurements, including the number, frequency, and amplitude of pressure peaks as well as the motility index (MI). Key Results-A total of 20 subjects were included (mean age 13.1 ± 3.4 years). No significant metabolite changes were observed at 10 min after the meal, whereas 16 amino acid and 22 lipid metabolites had significant (P < 0.005) postprandial changes, including decreases in methylhistamine, histamine, and GABA, by 60 min. Correlations were observed between normal and abnormal postprandial motility patterns and changes in specific metabolites, including glycerol, carnosine, alanine, asparagine, cytosine, choline, phosphocholine, thyroxine, and triiodothyronine. Interestingly, subjects without the normal postprandial increase in area under the curve (AUC), had markedly increased levels of kynurenic acid and adenosyl-homocysteine. Conclusions & Inferences-This is the first study to examine postprandial metabolic changes in children and also to correlate changes in specific metabolites with colonic motility. The results suggest possible metabolic pathways associated with motility and identify potential targets for the treatment of constipation.
Journal of anatomy, Jan 18, 2018
The enteric nervous system shares embryological, morphological, neurochemical, and functional fea... more The enteric nervous system shares embryological, morphological, neurochemical, and functional features with the central nervous system. In addition to neurons and glia, the CNS includes a third component, microglia, which are functionally and immunophenotypically similar to macrophages, but a similar cell type has not previously been identified in enteric ganglia. In this study we identify a population of macrophages in the enteric ganglia, intermingling with the neurons and glia. These intraganglionic macrophages (IMs) are highly ramified and express the hematopoietic marker CD45, major histocompatibility complex (MHC) class II antigen, and chB6, a marker specific for B cells and microglia in avians. These IMs do not express antigens typically associated with T cells or dendritic cells. The CD45 /ChB6 /MHCII signature supports a hematopoietic origin and this was confirmed using intestinal chimeras in GFP-transgenic chick embryos. The presence of green fluorescent protein positive (...
Infection and Immunity, 2017
Enteropathogenic Escherichia coli (EPEC) is a leading cause of severe intestinal disease and infa... more Enteropathogenic Escherichia coli (EPEC) is a leading cause of severe intestinal disease and infant mortality in developing countries. Virulence is mediated by a type three secretion system (T3SS), causing the hallmark attaching and effacing (AE) lesions and actin-rich pedestal formation beneath the infecting bacteria on the apical surface of enterocytes. EPEC is a human-specific pathogen whose pathogenesis cannot be studied in animal models. We therefore established an EPEC infection model in human gut xenografts in SCID mice and used it to study the role of T3SS in the pathogenesis of the disease. Following EPEC O127:H6 strain E2348/69 infection, T3SS-dependent AE lesions and pedestals were demonstrated in all infected xenografts. We report here the development of T3SS-dependent intestinal thrombotic microangiopathy (iTMA) and ischemic enteritis in ∼50% of infected human gut xenografts. Using species-specific CD31 immunostaining, we showed that iTMA was limited to the larger human...
Scientific reports, May 31, 2017
Mechanisms mediating adult enteric neurogenesis are largely unknown. Using inflammation-associate... more Mechanisms mediating adult enteric neurogenesis are largely unknown. Using inflammation-associated neurogenesis models and a transgenic approach, we aimed to understand the cell-source for new neurons in infectious and inflammatory colitis. Dextran sodium sulfate (DSS) and Citrobacter rodentium colitis (CC) was induced in adult mice and colonic neurons were quantified. Sox2GFP and PLP1GFP mice confirmed the cell-type specificity of these markers. Sox2CreER:YFP and PLP1creER:tdT mice were used to determine the fate of these cells after colitis. Sox2 expression was investigated in colonic neurons of human patients with Clostridium difficile or ulcerative colitis. Both DSS and CC led to increased colonic neurons. Following colitis in adult Sox2CreER:YFP mice, YFP initially expressed predominantly by glia becomes expressed by neurons following colitis, without observable DNA replication. Similarly in PLP1CreER:tdT mice, PLP1 cells that co-express S100b but not RET also give rise to neur...
Journal of anatomy, Jan 24, 2017
The development of the enteric nervous system (ENS) and intestinal smooth muscle occurs in a spat... more The development of the enteric nervous system (ENS) and intestinal smooth muscle occurs in a spatially and temporally correlated manner, but how they influence each other is unknown. In the developing mid-gut of the chick embryo, we find that α-smooth muscle actin expression, indicating early muscle differentiation, occurs after the arrival of migrating enteric neural crest-derived cells (ENCCs). In contrast, hindgut smooth muscle develops prior to ENCC arrival. Smooth muscle development is normal in experimentally aganglionic hindguts, suggesting that proper development and patterning of the muscle layers does not rely on the ENS. However, inhibiting early smooth muscle development severely disrupts ENS patterning without affecting ENCC proliferation or apoptosis. Our results demonstrate that early intestinal smooth muscle differentiation is required for patterning the developing ENS.
Pediatric Research, 2017
Basic Science Investigation nature publishing group Background: Enteric neural stem/progenitor ce... more Basic Science Investigation nature publishing group Background: Enteric neural stem/progenitor cells (ENSCs) offer an innovative approach to treating Hirschsprung disease (HSCR) and other enteric neuropathies. However, postnatalderived human ENSCs have not been thoroughly characterized and their behavior in the embryonic and postnatal intestinal environment is unknown. Methods: ENSCs were isolated from the intestines of 25 patients undergoing bowel resection, including 7 children with HSCR. Neuronal differentiation and proliferation of ENSCs from submucosal and myenteric plexuses from patients with and without HSCR were characterized. ENSC migration and differentiation were studied following transplantation into embryonic chick neural crest, embryonic chick hindgut, and postnatal mouse aganglionic colon. results: The proliferative and neurogenic potential of ENSCs from HSCR intestine is equivalent to that of non-HSCR controls. Similarly, no difference was observed between myenteric-and submucosal-derived ENSCs. Postnatal ENSCs transplanted to embryonic neural crest pathways and to aneural hindgut migrate normally and differentiate into appropriate neural crest-derived cell types. ENSCs in postnatal mouse aganglionic colon differentiate into neurons and glia both ex vivo and in vivo. conclusions: ENSCs isolated from the postnatal intestine of patients with and without HSCR can behave like embryonic neural crest-derived cells. These results support the feasibility of cell-based therapy for future treatment of neurointestinal disease.
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Papers by Allan Goldstein