Second generation knockout sickle mice: The effect of HbF

Mary E. Fabry, Sandra M. Suzuka, Rona S. Weinberg, Christine Lawrence, Stephen M. Factor, John G. Gilman, Frank Costantini, Ronald L. Nagel

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

Sickle transgenic mice expressing exclusively human globins are desirable for studying pathophysiology and testing gene therapy strategies, but they must have significant pathology and show evidence of amelioration by antisickling hemoglobins. Mice were generated that expressed exclusively human sickle hemoglobin with 3 levels of HbF using their previously described sickle constructs (cointegrated human miniLCRα2 and miniLCRβs [PNAS 89:12150, 1992]), mouse α- and β-globin-knockouts, and 3 different human γ-transgenes. It was found that, at all 3 levels of HbF expression, these mice have balanced chain synthesis, nearly normal mean corpuscular hemoglobin, and, in some cases, F cells. Mice with the least adult HbF expression were the most severe. Progressive increase in HbF from less than 3% to 20% to 40% correlated with progressive increase in hematocrit (22% to 34% to 40%) and progressive decrease in reticulocyte count (from 60% to 30% to 13%). Urine concentrating ability was normalized at high HbF, and tissue damage detected by histopathology and organ weight were ameliorated by increased HbF. The γ-transgene that produces intermediate levels of HbF was introduced into knockout sickle mice described by Pàszty and coworkers that express the miniLCRα1GγA γδβs transgene and have fetal but not adult expression of HbF. It was found that the level of HbF required to ameliorate low hematocrit and normalize urine concentrating defect was different for the miniLCRα2βs and miniLCRα1GγA γδβs mice. We conclude that knockout mice with the miniLCRα2βs transgene and postnatal expression of HbF have sufficiently faithful sickle pathology to serve as a platform for testing antisickling interventions.

Original languageEnglish (US)
Pages (from-to)410-418
Number of pages9
JournalBlood
Volume97
Issue number2
DOIs
StatePublished - Jan 15 2001
Externally publishedYes

Fingerprint

Globins
Pathology
Transgenes
Knockout Mice
Hemoglobins
Sickle Hemoglobin
Gene therapy
Testing
Hematocrit
Kidney Concentrating Ability
Tissue
Reticulocyte Count
Defects
Erythrocyte Indices
Organ Size
Genetic Therapy
Transgenic Mice
Urine

ASJC Scopus subject areas

  • Hematology

Cite this

Fabry, M. E., Suzuka, S. M., Weinberg, R. S., Lawrence, C., Factor, S. M., Gilman, J. G., ... Nagel, R. L. (2001). Second generation knockout sickle mice: The effect of HbF. Blood, 97(2), 410-418. https://doi.org/10.1182/blood.V97.2.410

Second generation knockout sickle mice : The effect of HbF. / Fabry, Mary E.; Suzuka, Sandra M.; Weinberg, Rona S.; Lawrence, Christine; Factor, Stephen M.; Gilman, John G.; Costantini, Frank; Nagel, Ronald L.

In: Blood, Vol. 97, No. 2, 15.01.2001, p. 410-418.

Research output: Contribution to journalArticle

Fabry, ME, Suzuka, SM, Weinberg, RS, Lawrence, C, Factor, SM, Gilman, JG, Costantini, F & Nagel, RL 2001, 'Second generation knockout sickle mice: The effect of HbF', Blood, vol. 97, no. 2, pp. 410-418. https://doi.org/10.1182/blood.V97.2.410
Fabry ME, Suzuka SM, Weinberg RS, Lawrence C, Factor SM, Gilman JG et al. Second generation knockout sickle mice: The effect of HbF. Blood. 2001 Jan 15;97(2):410-418. https://doi.org/10.1182/blood.V97.2.410
Fabry, Mary E. ; Suzuka, Sandra M. ; Weinberg, Rona S. ; Lawrence, Christine ; Factor, Stephen M. ; Gilman, John G. ; Costantini, Frank ; Nagel, Ronald L. / Second generation knockout sickle mice : The effect of HbF. In: Blood. 2001 ; Vol. 97, No. 2. pp. 410-418.
@article{b59ec0ca69ee4453b9c8d5812178cf3d,
title = "Second generation knockout sickle mice: The effect of HbF",
abstract = "Sickle transgenic mice expressing exclusively human globins are desirable for studying pathophysiology and testing gene therapy strategies, but they must have significant pathology and show evidence of amelioration by antisickling hemoglobins. Mice were generated that expressed exclusively human sickle hemoglobin with 3 levels of HbF using their previously described sickle constructs (cointegrated human miniLCRα2 and miniLCRβs [PNAS 89:12150, 1992]), mouse α- and β-globin-knockouts, and 3 different human γ-transgenes. It was found that, at all 3 levels of HbF expression, these mice have balanced chain synthesis, nearly normal mean corpuscular hemoglobin, and, in some cases, F cells. Mice with the least adult HbF expression were the most severe. Progressive increase in HbF from less than 3{\%} to 20{\%} to 40{\%} correlated with progressive increase in hematocrit (22{\%} to 34{\%} to 40{\%}) and progressive decrease in reticulocyte count (from 60{\%} to 30{\%} to 13{\%}). Urine concentrating ability was normalized at high HbF, and tissue damage detected by histopathology and organ weight were ameliorated by increased HbF. The γ-transgene that produces intermediate levels of HbF was introduced into knockout sickle mice described by P{\`a}szty and coworkers that express the miniLCRα1GγA γδβs transgene and have fetal but not adult expression of HbF. It was found that the level of HbF required to ameliorate low hematocrit and normalize urine concentrating defect was different for the miniLCRα2βs and miniLCRα1GγA γδβs mice. We conclude that knockout mice with the miniLCRα2βs transgene and postnatal expression of HbF have sufficiently faithful sickle pathology to serve as a platform for testing antisickling interventions.",
author = "Fabry, {Mary E.} and Suzuka, {Sandra M.} and Weinberg, {Rona S.} and Christine Lawrence and Factor, {Stephen M.} and Gilman, {John G.} and Frank Costantini and Nagel, {Ronald L.}",
year = "2001",
month = "1",
day = "15",
doi = "10.1182/blood.V97.2.410",
language = "English (US)",
volume = "97",
pages = "410--418",
journal = "Blood",
issn = "0006-4971",
publisher = "American Society of Hematology",
number = "2",

}

TY - JOUR

T1 - Second generation knockout sickle mice

T2 - The effect of HbF

AU - Fabry, Mary E.

AU - Suzuka, Sandra M.

AU - Weinberg, Rona S.

AU - Lawrence, Christine

AU - Factor, Stephen M.

AU - Gilman, John G.

AU - Costantini, Frank

AU - Nagel, Ronald L.

PY - 2001/1/15

Y1 - 2001/1/15

N2 - Sickle transgenic mice expressing exclusively human globins are desirable for studying pathophysiology and testing gene therapy strategies, but they must have significant pathology and show evidence of amelioration by antisickling hemoglobins. Mice were generated that expressed exclusively human sickle hemoglobin with 3 levels of HbF using their previously described sickle constructs (cointegrated human miniLCRα2 and miniLCRβs [PNAS 89:12150, 1992]), mouse α- and β-globin-knockouts, and 3 different human γ-transgenes. It was found that, at all 3 levels of HbF expression, these mice have balanced chain synthesis, nearly normal mean corpuscular hemoglobin, and, in some cases, F cells. Mice with the least adult HbF expression were the most severe. Progressive increase in HbF from less than 3% to 20% to 40% correlated with progressive increase in hematocrit (22% to 34% to 40%) and progressive decrease in reticulocyte count (from 60% to 30% to 13%). Urine concentrating ability was normalized at high HbF, and tissue damage detected by histopathology and organ weight were ameliorated by increased HbF. The γ-transgene that produces intermediate levels of HbF was introduced into knockout sickle mice described by Pàszty and coworkers that express the miniLCRα1GγA γδβs transgene and have fetal but not adult expression of HbF. It was found that the level of HbF required to ameliorate low hematocrit and normalize urine concentrating defect was different for the miniLCRα2βs and miniLCRα1GγA γδβs mice. We conclude that knockout mice with the miniLCRα2βs transgene and postnatal expression of HbF have sufficiently faithful sickle pathology to serve as a platform for testing antisickling interventions.

AB - Sickle transgenic mice expressing exclusively human globins are desirable for studying pathophysiology and testing gene therapy strategies, but they must have significant pathology and show evidence of amelioration by antisickling hemoglobins. Mice were generated that expressed exclusively human sickle hemoglobin with 3 levels of HbF using their previously described sickle constructs (cointegrated human miniLCRα2 and miniLCRβs [PNAS 89:12150, 1992]), mouse α- and β-globin-knockouts, and 3 different human γ-transgenes. It was found that, at all 3 levels of HbF expression, these mice have balanced chain synthesis, nearly normal mean corpuscular hemoglobin, and, in some cases, F cells. Mice with the least adult HbF expression were the most severe. Progressive increase in HbF from less than 3% to 20% to 40% correlated with progressive increase in hematocrit (22% to 34% to 40%) and progressive decrease in reticulocyte count (from 60% to 30% to 13%). Urine concentrating ability was normalized at high HbF, and tissue damage detected by histopathology and organ weight were ameliorated by increased HbF. The γ-transgene that produces intermediate levels of HbF was introduced into knockout sickle mice described by Pàszty and coworkers that express the miniLCRα1GγA γδβs transgene and have fetal but not adult expression of HbF. It was found that the level of HbF required to ameliorate low hematocrit and normalize urine concentrating defect was different for the miniLCRα2βs and miniLCRα1GγA γδβs mice. We conclude that knockout mice with the miniLCRα2βs transgene and postnatal expression of HbF have sufficiently faithful sickle pathology to serve as a platform for testing antisickling interventions.

UR - http://www.scopus.com/inward/record.url?scp=0035863541&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035863541&partnerID=8YFLogxK

U2 - 10.1182/blood.V97.2.410

DO - 10.1182/blood.V97.2.410

M3 - Article

C2 - 11154217

AN - SCOPUS:0035863541

VL - 97

SP - 410

EP - 418

JO - Blood

JF - Blood

SN - 0006-4971

IS - 2

ER -