34 Íîâîñòè êëåòî÷íûõ òåõíîëîãèé

34
Íîâîñòè êëåòî÷íûõ òåõíîëîãèé
7.Kondziolka D., Wechsler L., Goldstein S. et al. Transplantation of cultured
human neuronal cells for patients with stroke. Neurology 2000; 55: 565-9.
8.Nelson P.T., Kondziolka D., Wechsler L. et al. Clonal human (hNT) neuron
grafts for stroke therapy: neuropathology in a patient 27 months after
implantation. Am. J. Pathol. 2002; 160:1201-6.
9.Savitz S.I., Dinsmore J., Wu J. et al. Neurotransplantation of fetal porcine
cells in patients with basal ganglia infarcts: a preliminary safety and feasibility study.
Cerebrovasc. Dis. 2005; 20: 101-7.
10.Tateishi-Yuyama E., Matsubara H., Murohara T. et al. Therapeutic
Angiogenesis using Cell Transplantation (TACT) Study Investigators. Therapeutic
angiogenesis for patients with limb ischaemia by autologous transplantation of
bone-marrow cells: a pilot study and a randomised controlled trial. Lancet 2002;
360: 427-35.
11.Strauer B.E., Brehm M., Zeus T. et al. Repair of infracted myocardium by
autologous intracoronary mononuclear bone marrow cell transplantation in
humans. Circ. 2002;106: 1913-8.
12.Chen J., Li Y., Wang L. et al. Therapeutic benefit of intravenous
administration of bone marrow stromal cells after cerebral ischemia in rats. Stroke
2001; 32: 1005-11.
13.Li Y., Chen J., Chen X.G. et al. Human marrow stromal cell therapy for stroke
in rat: neurotrophins and functional recovery. Neurology 2002; 59: 514-23.
14.Zhao L.R., Duan W.M., Reyes M. et al. Human bone marrow stem cells
exhibit neural phenotypes and ameliorate neurological deficits after grafting into
the ischemic brain of rats. Exp. Neurol. 2002; 174: 11-20.
15.Chen J., Zhang Z.G., Li Y. et al. Intravenous administration of human bone
marrow stromal cells induces angiogenesis in the ischemic boundary zone after
stroke in rats. Circ. Res. 2003; 92: 692-9.
16.Massengale M., Wagers A.J., Vogel H., Weissman I.L. Hematopoietic cells
maintain hematopoietic fates upon entering the brain. J. Exp. Med. 2005; 201:
1579-89.
17.Chen X., Li Y., Wang L. et al. Ischemic rat brain extracts induce human
marrow stromal cell growth factor production. Neuropathology 2002; 22:
275-9.
18. Borlongan C.V., Hadman M., Sanberg C.D., Sanberg P.R. Central nervous
system entry of peripherally injected umbilical cord blood cells is not required for
neuroprotection in stroke. Stroke 2004; 35:2385-9.
Ïîäãîòîâèë Ä.Ñ. Ñòàíêîâ
ïî ìàòåðèàëàì Ann. Neurol. 2005; 57: 874-82
Èçó÷åíèå âûæèâàíèÿ è äèôôåðåíöèðîâêè àëëîãåííûõ
ôåòàëüíûõ êëåòîê, òðàíñïëàíòèðîâàííûõ â ãîëîâíîé ìîçã
ïàöèåíòàì ñ áîëåçíüþ Ïàðêèíñîíà - ðåçóëüòàòû
èññëåäîâàíèÿ àóòîïñèéíîãî ìàòåðèàëà
Ïåðâàÿ òðàíñïëàíòàöèÿ ôåòàëüíîé íåðâíîé òêàíè â ãîëîâíîé ìîçã ïàöèåíòàì ñ áîëåçíüþ Ïàðêèíñîíà (ÁÏ) áûëà
âûïîëíåíà â êëèíèêå â 1987 ãîäó ãðóïïîé Lindvall O. â Lund
University Hospital (Sweden) [1, 2]. C òåõ ïîð ñðàçó â íåñêîëüêèõ öåíòðàõ ìèðà áûëè íà÷àòû êëèíè÷åñêèå èñïûòàíèÿ ìåòîäà ñòåðåîòàêñè÷åñêîé íèãðîñòðèàëüíîé òðàíñïëàíòàöèè
ôåòàëüíîé íåðâíîé òêàíè. Åâðîïåéñêàÿ ïðîãðàììà NECTAR
(Network of European CNS Transplantation And Restoration),
ïðèçâàííàÿ îöåíèòü êëèíè÷åñêóþ ýôôåêòèâíîñòü ìåòîäà,
áûëà íà÷àòà â 1990 ãîäó è âêëþ÷àëà îêîëî 40 êëèíè÷åñêèõ
ãðóïï. Â ðåçóëüòàòå ýòèõ èñïûòàíèé áûëè ñäåëàíû âûâîäû,
÷òî ôåòàëüíûå àëëîãåííûå íèãðàëüíûå íåéðîíû ïðèæèâàþòñÿ áåç âûðàæåííîãî èììóííîãî îòâåòà, ðåèííåðâèðóþò
îêðóæàþùèé ñòðèàòóì ãîëîâíîãî ìîçãà õîçÿèíà (÷åëîâåêà),
âûçûâàþò îáðàçîâàíèå è âûáðîñ äîïàìèíà è îáåñïå÷èâàþò
óëó÷øåíèå ìîòîðíîé ôóíêöèè [2, 3, 6].
Îäíàêî, â äâóõ äâîéíûõ-ñëåïûõ, ïëàöåáî-êîíòðîëèðóåìûõ
(ïàöèåíòû áûëè ëîæíîîïåðèðîâàíû) èñïûòàíèÿõ, ñïîíñèðóåìûõ NIH â ÑØÀ, íå áûëî âûÿâëåíî çíà÷èìûõ ðàçëè÷èé â
óëó÷øåíèè ìîòîðíîé ôóíêöèè ìåæäó ãðóïïàìè è íåîæèäàííî
áûëè ïîëó÷åíû ïîáî÷íûå ýôôåêòû ó íåêîòîðûõ ïàöèåíòîâ
[4, 5]. Ïåðâûå ìîðôîëîãè÷åñêèå èññëåäîâàíèÿ, ïîäòâåðæäàþùèå äîëãîâðåìåííîå âûæèâàíèå ïåðåñàæåííûõ êëåòîê
è ÷àñòè÷íóþ ðåèííåðâàöèþ íèãðàñòðèàëüíîé îáëàñòè õîçÿèíà, áûëè âûïîëíåíû â 1995 ãîäó íà àóòîïñèéíîì ìàòåðèàëå ãîëîâíîãî ìîçãà ïàöèåíòîâ, ïîäâåðãøèõñÿ ïðîöåäóðå. Ýòè
æå äàííûå áûëè ïîäòâåðæäåíû â áîëåå ñîâðåìåííûõ èññëåäîâàíèÿõ Olanow C.W. [5] è Freed C.R. [4]. Âñå ýòè èññëåäîâàíèÿ àíàëèçèðîâàëè ñóäüáó ôðàãìåíòîâ òêàíåâîãî òðàíñïëàíòàòà, íî íå ñóñïåíçèè íåéðîêëåòîê, ïîäâåðãøèõñÿ
ïðåäâàðèòåëüíîé ýíçèìàòè÷åñêîé îáðàáîòêå è êóëüòèâèðîâàíèþ. Îäíàêî, èìåííî ïî ïðè÷èíå ïðèìåíåíèÿ ðàçëè÷íûõ
âèäîâ òðàíñïëàíòàòîâ ðÿä èññëåäîâàòåëåé âèäèò ðàçíèöó,
ïîëó÷åííóþ â êëèíè÷åñêèõ èññëåäîâàíèÿõ íåçàâèñèìûõ
ãðóïï. Òàêèì îáðàçîì, èñïîëüçîâàíèå òêàíåâîãî èëè
Êëåòî÷íàÿ òðàíñïëàíòîëîãèÿ è òêàíåâàÿ èíæåíåðèÿ ¹ 2, 2005
ñóñïåíçèîííîãî òðàíñïëàíòàòà îñòà¸òñÿ íåðåø¸ííûì âîïðîñîì êëèíè÷åñêîé íåéðîòðàíñïëàíòàöèè.
 æóðíàëå Brain îïóáëèêîâàíû ðåçóëüòàòû ïàòîìîðôîëîãè÷åñêîãî èññëåäîâàíèÿ àóòîïñèéíîãî ìàòåðèàëà ãîëîâíîãî
ìîçãà ïàöèåíòîâ ñ ÁÏ, ïîëó÷èâøèõ ñóñïåíçèîííûé íåéðîòðàíñïëàíòàò áîëåå 3-õ ëåò íàçàä.
Ïðîäóêöèÿ äîôàìèíà
ïî äàííûì F-18-DOPA
ïîçèòðîííî-ýìèññèîííîé
òîìîãðàôèè ãîëîâíîãî ìîçãà.
Ñëåâà - äî ëå÷åíèÿ, ñïðàâà
÷åðåç 3 ãîäà ïîñëå
òðàíñïëàíòàöèè.
Èç Brain 2005; 128:
1498-510
Àíàëèçó ïîäâåðãëè òêàíü ãîëîâíîãî ìîçãà 2-õ ðåöèïèåíòîâ, ïîëó÷èâøèõ òðàíñïëàíòàöèè ôåòàëüíûõ êëåòîê è
óìåðøèõ îò îñëîæíåíèé îñòðîãî èíôàðêòà ìèîêàðäà ÷åðåç
3 ãîäà 8 ìåñÿöåâ è 4 ãîäà 4 ìåñÿöà ïîñëå âìåøàòåëüñòâà.
Âàæíî, ÷òî â ýòîì èññëåäîâàíèè ïåðåñàæèâàëè ñóñïåíçèþ
êëåòîê, âûäåëåííóþ ýíçèìàòè÷åñêè (èç òêàíè ñðåäíåãî ìîçãà
6-9 íåäåëüíûõ ýìáðèîíîâ), ïðåäâàðèòåëüíî êóëüòèâèðîâàííûõ â ñðåäå ñ ôàêòîðàìè ðîñòà (GDNF). Òðàíñïëàíòàöèè âûïîëíÿëè ñòåðåîòàêñè÷åñêè áèëàòåðàëüíî â íåñêîëüêî òî÷åê
ïî ðàçíûì òðàåêòîðèÿì â îáëàñòè ñòðèàòóìà (ïóòàìåí) è ÷¸ðíîé ñóáñòàíöèè. Ïåðâûé ïàöèåíò ïîëó÷èë 3,2 ìëí êëåòîê,
âòîðîé - 4,8 ìëí (îáà - îò 3-õ ýìáðèîíîâ). Ïîñëå òðàíñïëàíòàöèè îáà ïàöèåíòà ïðîäåìîíñòðèðîâàëè êëèíè÷åñêîå óëó÷øåíèå è ïðèçíàêè ñèíòåçà äîïàìèíà â îáëàñòè òðàíñïëàíòàöèè. Àâòîðû ïîäñ÷èòàëè êîëè÷åñòâî äîïàìèí-íåéðîíîâ.
Áîëüøàÿ ÷àñòü äîíîðñêèõ êëåòîê (40-60%) èìåëà ôåíîòèï
35
Human study
äîïàìèí-íåéðîíîâ äâèãàòåëüíîé çîíû ñòðèàòóìà íîðìàëüíîãî ìîçãà ÷åëîâåêà (Calbidin-/Girk2+). Äîïàìèí-íåéðîíû
òðàíñïëàíòàòà íàõîäèëè â êàæäîì êàíàëå ââåäåíèÿ êëåòîê è
â âèäå ñêîïëåíèé - â ìåñòàõ ãðàôòèíãà. Îáùåå êîëè÷åñòâî
òàêèõ íåéðîíîâ ñîñòàâëÿëî îêîëî 120-200 òûñÿ÷. Íåñìîòðÿ
íà òî, ÷òî ïàöèåíòû ïðèíèìàëè èììóíîñóïðåññîðû (öèêëîñïîðèí À) â òå÷åíèå ïîëóãîäà ïîñëå òðàíñïëàíòàöèè, áûëè âûÿâëåíû ëèøü ïðèçíàêè íåáîëüøîãî ñêîïëåíèÿ (ïîÿâëåíèÿ òàê
íàçûâàåìûõ «ðóáöîâ») CD45+ êëåòîê àêòèâèðîâàííîé ìèêðîãëèè (GFAP+) è åäèíè÷íûå ìàêðîôàãè. Ýòè äàííûå ïðîòèâîðå÷àò ìîðôîëîãè÷åñêèì äàííûì, ïîëó÷åííûì â èññëåäîâàíèè
Freed C.R. [4], ãäå áûëà âûÿâëåíà âûðàæåííàÿ ìèêðîãëèàëüíàÿ
ðåàêöèÿ è ïîâûøåííàÿ ýêñïðåññèÿ MHC-II (ãåíîâ ãèñòîñîâìåñòèìîñòè) ïðè òðàíñïëàíòàöèè ôðàãìåíòîâ òêàíè.
Òàêèì îáðàçîì, â èññëåäîâàíèè áûëî ïîêàçàíî, ÷òî ôåòàëüíûå íåéðîíû, òðàíñïëàíòèðîâàííûå â ãîëîâíîé ìîçã ïàöèåíòîâ ñ áîëåçíüþ Ïàðêèíñîíà, äëèòåëüíî âûæèâàþò (áîëåå 3-4 ëåò). Êëåòêè ïðîäóöèðóþò òåðàïåâòè÷åñêèé óðîâåíü
äîïàìèíà è ðåèííåðâèðóþò ïóòàìåí õîçÿèíà áåç ïîáî÷íûõ
ýôôåêòîâ. Àâòîðû óêàçûâàþò, ÷òî ïîñëå òðàíñïëàíòàöèè â
îáëàñòü ïóòàìåíà âûæèâàåìîñòü äîïàìèí+ êëåòîê ñîñòàâèëà 15-30%, à òðàíñïëàíòàòà îáëàñòè ÷¸ðíîé ñóáñòàíöèè 4-8%. Õîðîøèé ýôôåêò òðàíñïëàíòàöèè àâòîðû îáúÿñíÿþò
ïðèìåíåíèåì ìåòîäà ïðåêóëüòèâèðîâàíèÿ ñóñïåíçèè íåéðîíîâ â ñðåäå ñ GDNF è ââåäåíèÿ èìåííî ñóñïåíçèè êëåòîê,
ËÈÒÅÐÀÒÓÐÀ:
1. Lindvall O., Brundin P., Widner H. et al. Grafts of fetal dopamine neurons survive
and improve motor function in Parkinson's disease. Science 1990; 247: 574-7.
2. Winkler C., Kirik D., Bjorklund A. Cell transplantation in Parkinson's disease:
how can we make it work? Trends Neurosci. 2005; 28: 86-92.
3. Dunnett S.B., Bjorklund A., Lindvall O. Cell therapy in Parkinson's disease stop or go? Nat Rev. Neurosci. 2001; 2: 365-9.
4. Freed C.R., Greene P.E., Breeze R.E. et al. Transplantation of embryonic
à íå ôðàãìåíòîâ òêàíè, êàê ýòî áûëî âûïîëíåíî â äâóõ ïðåäûäóùèõ èññëåäîâàíèÿõ. Êðîìå òîãî, â ýòîì èññëåäîâàíèè
âïåðâûå áûëî ïðîàíàëèçèðîâàíî ñîñòîÿíèå òðàíñïëàíòàòà
ïîñëå ãðàôòèíãà â îáëàñòü ÷¸ðíîé ñóáñòàíöèè. Îáùåå ÷èñëî
âûæèâøèõ äîïàìèí+ íåéðîíîâ áûëî çíà÷èòåëüíî âûøå, ÷åì
èõ ïðèìåðíîå êîëè÷åñòâî â òâ¸ðäûõ òðàíñïëàíòàòàõ, óêàçàííîå àâòîðàìè [4, 5]. Àâòîðû èññëåäîâàíèÿ óêàçûâàþò, ÷òî
ïðèìåíåíèå ïîäîáíîãî ìåòîäà çàìåñòèòåëüíîé êëåòî÷íîé
òðàíñïëàíòàöèè íå èìååò ïîáî÷íûõ ýôôåêòîâ è ìîæåò áûòü
òåðàïåâòè÷åñêè âûãîäíûì äàæå ó ïàöèåíòîâ íà äàëåêî çàøåäøèõ ñòàäèÿõ çàáîëåâàíèÿ ñ àíàìíåçîì áîëåå 10 ëåò.
Íàïðèìåð, ó îäíîãî ïàöèåíòà áûëî äîñòèãíóòî äîëãîâðåìåííîå âîññòàíîâëåíèå ñèíòåçà äîïàìèíà ñëåâà íà 65% è ñïðàâà - íà 88% îò íîðìàëüíîãî óðîâíÿ.
Ïîñëå èñïûòàíèé ìåòîäà íåéðîòðàíñïëàíòàöèè äëÿ ëå÷åíèÿ áîëåçíè Ïàðêèíñîíà Olanow [5] è Freed [4], åãî êëèíè÷åñêàÿ ñóäüáà áûëà ïîñòàâëåíà ïîä ñîìíåíèå. Îäíàêî, òùàòåëüíûå ìîðôîëîãè÷åñêèå èññëåäîâàíèÿ ïîñòìîðòàëüíûõ
òðàíñïëàíòàòîâ â ãîëîâíîì ìîçãå ðåöèïèåíòîâ ïîêàçûâàþò,
÷òî ìåòîä «ðàáîòàåò» è ìîæåò áûòü êëèíè÷åñêè âûãîäíûì
äëÿ áîëüøèíñòâà ïàöèåíòîâ. Òàêèå èññëåäîâàíèÿ óêàçûâàþò êëèíèöèñòàì, êàêîé èìåííî ïðîòîêîë íåéðîòðàíñïëàíòàöèè îïòèìàëåí è ìîæåò ñëóæèòü ìåòîäîì âûáîðà. Äèñêóññèÿ
î êëèíè÷åñêîé ñóäüáå è ïåðñïåêòèâàõ ìåòîäà ïðåäñòàâëåíà
â íåäàâíåì îáçîðå [2].
dopamine neurons for severe Parkinson's disease. N. Engl. J. Med. 2001; 344: 710-9.
5. Olanow C.W., Goetz C.G., Kordower J.H. et al. A double-blind controlled
trial of bilateral fetal nigral transplantation in Parkinson's disease. Ann. Neurol.
2003; 54: 403-14.
6. Kordower J.H., Freeman T.B., Snow B.J. et al. Neuropathological evidence
of graft survival and striatal reinnervation after the transplantation of fetal
mesencephalic tissue in a patient with Parkinson's disease. N. Engl. J. Med. 1995;
332: 1118-24.
Ïîäãîòîâèë À.Â. Áåðñåíåâ
ïî ìàòåðèàëàì Brain 2005; 128: 1498-510;
doi:10.1093/brain/awh510
Òðàíñïëàíòàöèÿ êëåòîê êîñòíîãî ìîçãà ïàöèåíòàì
ñ òðàâìîé ñïèííîãî ìîçãà - ðåçóëüòàòû ïèëîòíîãî
êëèíè÷åñêîãî èññëåäîâàíèÿ â Þæíîé Êîðåå
Òðàâìà ïîçâîíî÷íèêà, îñëîæíåííàÿ ïîâðåæäåíèåì
ñïèííîãî ìîçãà â âèäå åãî êîìïðåññèè, ðàçìîçæåíèÿ, ÷àñòè÷íîãî èëè ïîëíîãî ðàçðûâà, ÿâëÿåòñÿ îäíîé èç àêòóàëüíûõ
ìåäèêî-ñîöèàëüíûõ ïðîáëåì ñîâðåìåííîé ìåäèöèíû, ïîñêîëüêó âåä¸ò ê ãëóáîêîé èíâàëèäèçàöèè ïîñòðàäàâøèõ.
Ïîñëåäñòâèÿ òðàâìû ñïèííîãî ìîçãà (ÒÑÌ) ÷àùå âñåãî
ïðîÿâëÿþòñÿ âÿëûì èëè ñïàñòè÷åñêèì ïàðàëè÷îì, â ëó÷øåì ñëó÷àå - ïàðåçîì êîíå÷íîñòåé è äèñôóíêöèåé òàçîâûõ
îðãàíîâ.  íàñòîÿùåå âðåìÿ íå ñóùåñòâóåò äåéñòâèòåëüíî ýôôåêòèâíûõ ìåòîäîâ ëå÷åíèÿ òðàâìàòè÷åñêîãî ïîâðåæäåíèÿ ñïèííîãî ìîçãà, îñîáåííî êîãäà ïîñëå òðàâìû
ïðîøëè ìåñÿöû è ãîäû [1].
 ïîñëåäíèå ãîäû áûëè èñïûòàíû ðàçëè÷íûå ìåòîäû
êëåòî÷íîé íåéðîòðàíñïëàíòàöèè äëÿ êîððåêöèè ÒÑÌ â êëèíèêå [2]. Äëÿ ýòîãî ïðèìåíÿëè òðàíñïëàíòàöèþ ïîääåðæèâàþùèõ êëåòîê «îáîíÿòåëüíîãî ýïèòåëèÿ» [3], àêòèâèðîâàííûõ
ìàêðîôàãîâ [4], ôåòàëüíûõ íåéðîíîâ [5] è êëåòîê êîñòíîãî
ìîçãà [6].
 ïðîøëîì ãîäó íà åæåãîäíîé êîíôåðåíöèè American
Society for Neurotransplantation and Repair áûë ïðåäñòàâëåí
ïåðâûé äîêëàä ïî ðåçóëüòàòàì àóòîëîãè÷íîé òðàíñïëàíòàöèè êëåòîê êîñòíîãî ìîçãà äëÿ ëå÷åíèÿ ÒÑÌ â Motol Hospital
(Prague) [6]. Îò 10 äî 147 ìëí CD34+ êëåòîê êîñòíîãî ìîçãà
ââîäèëè â ïîçâîíî÷íûå àðòåðèè 9-òè ïàöèåíòàì ñ ðàçëè÷íûìè óðîâíÿìè ïîâðåæäåíèÿ. Ó 6-òè èç 9-òè ïàöèåíòîâ áûëî
îòìå÷åíî óëó÷øåíèå ÷åðåç 3-6 ìåñÿöåâ ïîñëå òðàíñïëàíòàöèè êëåòîê [2, 6].
 íåäàâíåì íîìåðå æóðíàëà Tissue Engineering îïóáëèêîâàíû ðåçóëüòàòû êëèíè÷åñêîãî èññëåäîâàíèÿ ïî ýôôåêòèâíîñòè òðàíñïëàíòàöèè êëåòîê êîñòíîãî ìîçãà â îáëàñòü
ïîâðåæäåíèÿ ñïèííîãî ìîçãà â ñî÷åòàíèè ñ òåðàïèåé ãðàíóëîöèòàðíî-ìàêðîôàãàëüíûì êîëîíèåñòèìóëèðóþùèì
Êëåòî÷íàÿ òðàíñïëàíòîëîãèÿ è òêàíåâàÿ èíæåíåðèÿ ¹ 2, 2005