Experimental Fetal Medicine

There are two themes to this work:

1. Fetal Circulating Cell Biology

Composition:

Professor Nicholas Fisk
Dr Pascale Guillot
Dr Jerry Chan
Dr Keelin O’Donoghue
Dr Sailesh Kumar
Dr Hitoshi Kurata

Principle aim:

The investigation of human fetal mesenchymal stem cell biology as applied to fetal development, fetomaternal cell traffic and fetal therapy.

Important findings in the last few years:

Isolation and characterisation of mesenchymal stem cells (MSC) in first trimester blood, liver and bone marrow
Differentiation of human fetal MSC into mesenchymal lineages in vitro and in vivo
Optimisation of myogenesis in vitro and in vivo with galectin 1
Lentiviral transduction has no effect on long-term stem cell properties
Demonstration that fetal MSC normally persist in maternal bone and bone marrow for decades after pregnancy

Plan of research for near future:

Understanding the molecular regulation of hfMSC multipotency and differentiation.
Determine hfMSC fate in parenchymal organs and bone marrow after intrauterine transplantation
Studying the potential of first trimester fetal MSC as vehicles for fetal cellular therapy and autologous gene delivery in experimental models of debilitating bone and muscle diseases
Investigating in vitro how fetal MSC support haemopoiesis
Develping safe ultrasound guided and endoscopic clinical techniques for hfMSC harvest

Links / collaborations:

Prof Irene Roberts & Dr Josu de la Fuente, Department of Haematology, Imperial College London (haemopoietic progenitors, cotransplantation)

Dr Jenny Morgan and Professor Terry Partridge, Muscle Biology Group, MRC Clinical Sciences Centre (myogenic differentiation, muscular dystrophy models)

Dr Huseyin Mehmet and Dr N Kennea, Institute of Reproductive & Developmental Biology, Imperial College London (neural stem cells, models of perinatal brain injury)

Professor Dame Julia Polak and Dr Ann Bishop, Tissue Engineering Centre, Imperial college (tissue engineering, osteoegenic dfferentiation, non viral transduction)

Drs Mike Themis and Simon Waddington, Gene Therapy Group, Imperial College London (viral integration, fetal transplantation models)

Professor Diana Watt, Brighton* and Sussex Medical School, (galectin in myogenesis)

Dr Jacquie Kerr, Laboratory of the Government Chemist (LGC) (valid analytical measurement, free fetal DNA in maternal blood)

Specific projects:

Human fetal mesenchymal stem cell fate after intrauterine transplantation (with Professor M Alison, Dr H Mehemt, and Dr M Themis) funded by a MRC Strategic Stem Cell Grant).

Intrauterine human fetal mesenchymal stem cell transportation for osteogenesis imperfecta (with Professor J Polak) funded by Action Medical Research.

Special non-invasive advances in fetal and neonatal evalutions network (SAFEN) (with Dr S Hahn and 53 others) funded by the European Community.

Fetal stem cell microchimerism in post-reproductive females (with Professor I Roberts), funded by Hammersmith Hospitals Trustees' Research Committee.

High-end multipurpose ultrasound for pre-clinical experimental research (with Professor M Blomley), funded by a Medical Research Council Joint Research Equipment Initiative.

Fetal mesenchymal stem cell as vectors for ex vivo autologous fetal gene therapy for musculoskeletal disorders (Dr Jerry Chan, Dr Sailesh Kumar) funded by Hammersmith Hospitals Trust.

Fetal mesenchymal stem cells as potential therapy for newborn brain injury (Dr N Kennea) funded by a Research Training Fellowship from the Wellcome Trust.

The role of mesenchymal stem cells in fetal haemopoiesis. (Dr de la Fuente) funded by a Research Training Fellowship from the Medical Research Council.

Recent key publications:

O'Donoghue K, Chan J, de la Fuente J, Kennea N, Sandison A, Anderson JR, Roberts IAG, Fisk NM: Microchimerism in female bone marrow and bone decades after mesenchymal stem cell trafficking in pregnancy. Lancet 364: 179-82, 2004.

Chan J, O'Donoghue K, de la Fuente J, Roberts IA, Kumar S, Morgan JE, fisk NM: Human fetal mesenchymal stem cells as vehicles for gene delivery. Stem Cells (In press).

O'Donoghue K, Choolani M, Chan J, de la Fuente J, Kumar S, Campagnoli C, Bennett PR, Roberts IAG, Fisk NM: Identification of fetal mesenchymal stem cells in maternal blood: implications for non-invasive prenatal diagnosis. Mol Hum Reprod 9, 497-502, 2003.

Choolani M, O'Donoghue K, Talbert D, Kumar S, Roberts I, Letsky E, Bennett PR, Fisk NM: Characterisations of first trimester fetal erythroblasts for non-invasive prenatal diagnosis. Mol Hum Reprod 9:227-35, 2003

Tocci A, Roberts IR, Kumar S, Bennett P, Fisk NM: CD34⁺cells from first trimester fetal blood are enriched in haemopoietic primitive progenitors. Am J Obstet Gynecol 188:1002-10, 2003

Campagnoli C, Bellantuono I, Kumar S, Fairbairn LJ, Roberts I, Fisk NM: High transduction efficiency of circulating fetal mesenchymal stem cells: potential targets for in utero ex vivo gene therapy. Br J Obstet Gynecol 109: 952-4, 2002.

Campagnoli C, Roberts IAG, Kumar S, Choolani M, Bennett PR, Letsky E, Fisk NM: Expandability of haemopoietic progenitors in first trimester fetal and maternal blood: implications for non-invasive prenatal diagnosis. Prenat Diagn 22: 463-9, 2002.

Choolani M, O’Donnell H, Campagnoli C, Kumar S, Roberts I, Bennett P, Fisk NM: Simultaneous fetal cell identification and diagnosis by epsilon globin chain immunophenotyping and chromosomal FISH. Blood, 98: 554-7, 2001.

Campagnoli C, Roberts IAG, Kumar S, Bennett PR, Bellantuono I, Fisk NM: Identification of mesenchymal stem/progenitor cells in human first-trimester fetal blood, liver, and bone marrow. Blood. 98:2396-2402, 2001.

Campagnoli C, Fisk NM, Bennett, P, Overton T, Roberts I: Circulating multipotent haemopoietic progenitors in first trimester fetal blood. Blood 95: 1967-72, 2000.

Male MSC among bone marrow cells in a woman 50
years after bearing a son (left) and persistent male
cell in post-reproductive female bone (right)

Galectin-1 induced muscle formation
(desmin stain Day 12) of human
fetal mesenchymal stem cells

GPF +ve human fetal mesenchymal stem
cells in fetal heart following intrauterine
transplantation

2. Monochorionic Placentation
Composition:

Prof Nicholas Fisk
Dr Helena Gardiner
Dr David Talbert
Dr Venu Jain
Dr Paula Galea
Dr Olivia Barigye
Dr Ling Wee
Dr Ruwan Wimalasundera

Principle aim:

The investigation of the placental and circulatory pathophysiology of intertwin transfusion syndromes in monochorionic twins.

Important achievements in the last few years:

Delineation of the placental vascular anastomotic basis of twin twin transfusion syndrome (TTTS)
Demonstration of discordant vascular programming in utero in identical twins
Development of Doppler techniques for studying arterio arterial and arteriovenous anastomoses in vivo
Development of clinic test to predict TTTS and its outcome, and to select optimal therapy.

Plan of research for near future:

Determine factors regulating anastomotic formation and blood flow
Characterise discordant renin angiotensin activation in TTTS and its effect on the fetoplacental vasculature
Characterization of structure and function of the shared cotyledon
Development of imaging methods for delineating vascular anatomy in monochorionic twins in vivo and quantitating intertwin transfusion

Links / collaborations:

Dr Mark Sullivan, Institute of Reproductive & Developmental Biology, Imperial College London (trophoblast function)
Professor Mark Kilby University of Birmingham (RAS activation)
Dr Frances Cowan, Dr Mary Rutherford, Imperial College London, Dept Paediatrics and MRC Clinical Sciences Centre (fetal and neonatal MRI)
Dr Kim Parker, Department of Physics, Imperial College London (anastomotic interference patterns, vascular casts)
Dr Ken Moise, Chapel Hill, North Carolina (multicentre septostomy trial)
Dr Mark Smith, Qinetiq (fetal ECG recording)
Dr Rob Cincotta, University of Queensland, Australia (experimental modelling of twin-twin transfusion syndrome)

Specific projects:

Discordant fetal renin-angiotensin system activation and fetoplacental vascular reactivity in twin-twin transfusion syndrome (Dr Galea), funded by the Richard & Jack Wiseman Trust.

Vascular pathophysiology of intertwin transfusion syndromes (Dr Wee), funded by the Richard & Jack Wiseman Trust.

Recent key publications:

Wee LY, Taylor MJO, Watkins N, Franke V, Parker K, Fisk NM: Characterisation of deep arterio-venous anastomoses within monochorionic placentae by vascular casting. Placenta (in press)

Tan TYT, Taylor MJO, Wee LY, Vanderheyden T, Wimalasundera R, Fisk NM: Doppler for artery-artery anastomosis predicts stage independent survival in twin-twin transfusion. Obstet Gynecol 103:1174-80, 2004.

Denbow ML, Taylor MJO, Cox P, Fisk NM: Derivation of rate of interfetal transfusion by analysis of arterio-arterial anastomotic waveform pattern in utero. Placenta 25: 664-70, 2004.

Wee LY, Taylor MJO, Vanderheyden T, Wimalasundera R, Gardiner HM, Fisk NM: Reversal of twin-twin transfusion syndrome: frequency, vascular anatomy, associated anomalies and outcome. Prenat Diagn 24:104-110, 2004

Tan TYT, Denbow ML, Cox PM, Talbert D, Fisk NM: Occlusion of arterio-arterial anastomosis manifesting as acute twin-twin transfusion syndrome. Placenta 25, 238-42, 2004

Taylor MJO, Talbert D, Fisk NM: Pseudo-arterio-arterial anastomosis in twin-twin transfusion syndrome. Placenta (in press)

Wee LY, Taylor MJO, Vanderheyden T, Talbert D, Fisk NM: Transmitted arterio-arterial anastomosis waveforms causing cyclically intermittent absent/reversed end-diastolic umbilical artery flow in monochorionic twins. Placenta 24, 772-8, 2003

Taylor MJO, Smith MJ, Thomas M, Green A, Cheng, Oseku-Afful S, Wee L, Fisk NM, Gardiner HM: Non-invasive fetal electrocardiography in singleton and multiple pregnancies. Br J Obstet Gynaecol 110, 668-78, 2003.

Franke V, Sherwin S, Parker K, Watkins N, Ling W, Fisk NM: Time domain computational modelling of 1 D arterial networks in the placenta. ESAIM: M²AN 37, 557-80, 2003.

Taylor MJO, Govender L, Jolly M, Wee L, Fisk NM: Validation of the Quintero staging system for twin-twin transfusion syndrome. Obstet Gynecol 100:1257-65, 2002.

Gardiner HM, Taylor MJO, Karatza A, Vanderheyden T, Huber A, Greenwald SE, Fisk NM, Hecher K. Twin-twin transfusion syndrome: the influence of intrauterine laser photocoagulation on arterial distensibility in childhood. Circulation 107:1906-11, 2003.

Karatza AA, Wolfenden JL, Taylor MJO, Wee L, Fisk NM, Gardiner HG: The influence of twin-twin transfusion syndrome on fetal cardiovascular structure and function: prospective case-control study of 136 monochorionic twin pregnancies. Heart 88: 271-7, 2002.

Tanawattanacharoen S, Taylor MJO, Jolly M, Letsky L, Cox P, Fisk NM: Intrauterine rescue transfusion in monochorionic multiple pregnancies with single intrauterine death. Prenat Diagn 21: 274-8, 2001.

Sebire NJ, Talbert D, Fisk NM: Twin-to-twin transfusion syndrome results from asymmetry in the progressive reduction of an initially symmetrical, large number of bi-directional arterio-venous connections formed during the embryonic unification of placental and fetal vessels. Placenta, 22: 383-91, 2001.

Mari G, Roberts A, Kovanci E, Detti L, Deter RL, Fisk NM: Perinatal morbidity and mortality in severe twin-twin transfusion syndrome: results of the International Amnioreduction Registry. Am J Obstet Gynecol, 185:708-15., 2001.

Welsh AW, Humphries K, Cosgrove D, Taylor MJO, Fisk NM: Development of freehand three dimensional power Dopper ultrasound and application to fetoplacental vasculature. Ultrasound in Medicine & Biology, 27:1161-70., 2001.

Jolly M, Taylor M, Govender L, Fisk NM: Interstitial laser therapy for twin reversed arterial perfusion sequence. BJOG 108, 1098—1102, 2001.

Welsh AW, Taylor MJO, Cosgrove D, Fisk NM: Freehand three -dimensional Doppler demonstration of monochorionic vascular anastomoses in vivo: a preliminary report. Ultrasound Obstet Gynecol 18:317-34, 2001.

Taylor MJO, Shalev E, Tanawattanacharoen S, Jolly M, Kumar S, Weiner E, Denbow ML, Cox PM, Fisk NM: Ultrasound guided umbilical cord occlusion using bipolar diathermy for Stage III/IV twin-twin transfusion syndrome. Prenat Diagn 22: 70-6, 2001.

Denbow ML, Rivens IH, Rowland IJ, Fisk NM, ter Haar GR, Leach MO: Pre-clinical development of non-invasive vascular occlusion using focused ultrasound surgery for fetal therapy. Am J Obstet Gynecol 182: 387-92, 2000.

Denbow ML, Cox P, Taylor M, Hammal DM, Fisk NM: Placental angioarchitecture in monochorionic twin pregnancies: relationship to fetal growth, feto-fetal transfusion syndrome, and pregnancy outcome. Am J Obstet Gynecol 182: 417-26, 2000.

Cheung YF, Taylor MJO, Fisk NM, Redington AN, Gardiner HM: Fetal origins of reduced arterial distensibility in the donor twin in twin-twin transfusion syndrome. Lancet 355:1157-8, 2000.

Denbow ML, Welsh A, Taylor MJ, Blomley MJK, Cosgrove DO, Fisk NM: Twin fetuses: intravascular microbubble US contrast administration in twin fetuses - early experience. Radiol 214: 724-8, 2000.

Denbow ML, Eckersly R, Taylor M, Welsh A, Carter R, Fisk NM: Ex vivo delineation of placental angioarchitecture with the microbubble contrast agent LevovistTM. Am J Obstet Gynecol 182: 966-71, 2000.

Taylor MJO, Denbow ML, Tanawattanacharoen S, Gannon C, Cox PM, Fisk NM: Doppler detection of arterio-arterial anastomoses in monochorionic twins: feasibility and clinical application. Hum Reprod 15: 1632-36, 2000.

Taylor MJO, Denbow ML, Duncan KR, Overton TG, Fisk NM: Antenatal factors at diagnosis predictive of outcome in severe twin-twin transfusion syndrome. Am J Obstet Gynecol 183: 1023-8, 2000.

Taylor MJO, Farquharson D, Cox P, Fisk NM: Identification of arterio-venous anastomoses in vivo in monochorionic twin pregnancies: preliminary report. Ultrasound Obstet Gynecol 16: 218-22, 2000.

Fisk NM, Howard C, Ware M, Bennett PR: X-chromosome inactivation patterns do not implicate asymmetric splitting of the inner cell mass in the aetiology of twin-twin transfusion syndrome. Mol Hum Reprod 5: 52-6, 1999.

Overton TG, Denbow ML, Duncan KR, Fisk NM: First-trimester cord entanglement in monochorionic twins. Ultrasound Obstet Gynecol 13: 140-2, 1999.

Denbow ML, Overton TG, Duncan KR, Cox PM, Fisk NM: High failure rate of umbilical vessel occlusion by ultrasound guided injection of absolute alcohol or enbucrilate gel. Prenat Diagn 19: 527-32, 1999.

Bajoria R, Sullivan M, Fisk NM: Endothelin levels in monochorionic twins with severe twin-twin transfusion syndrome. Hum Reprod 14: 1614-8, 1999.

Vascular cast of monochorionic placenta showing
a deep AV anastomosis (arrow) hidden beneath
the chorionic plate

A functional arterio-arterial anastomosis in vivo
detected with colour Doppler, showing characteristic
periodicity reflecting the net difference in fetal
heart rates