BENEFICIAL EFFECTS OF SUBCUTANEOUSLY INJECTED HUMAN UMBILICAL CORD STEM CELLS ON CEREBRAL PALSY AND TRAUMATIC BRAIN INJURY IN CHILDREN AND A POSITED MECHANISM (ANTHONY G. PAYNE)
BACKGROUND

Stem-cell laden human cord blood has proven of clear benefit in a diverse range of diseases includ- ing but not limited to leukemia [1], breast cancer [2], aplastic anemia [3], Fanconi’s anemia [4], and various immune disorders. CD34+ progenitor cells immunomagnetically separated from cord blood and introduced into animal models of various neu- rological conditions such as ALS, Alzheimer’s, Huntington’s, and stroke has demonstrated efficacy in terms of effecting notable improvement [5-9]. Laboratory research has shown that cord blood progenitor cells can be differentiated into os- teoblasts, chondroblasts, adipocytes, and hemato- poietic and neural cells including astrocytes and neurons [10]. hUCSC implanted into intact adult rat brains revealed that "human Tau-positive cells per- sisted for up to 3 months and showed migratory activity and a typical neuron-like morphology" [11]. "In vivo differentiation of hUCSC along mesodermal and endodermal pathways was demon- strated in animal models" [11]. In addition, human umbilical cord stem cells injected into the tail vein of rodents were subsequently found in the brains of the animals, thus providing proof these cells reach and penetrate the blood brain barrier [12].
In Mexico, research physician Fernando Rami- rez carried out a pilot study in which he treated 8 children with cerebral palsy with 1.5-2 million CD34+/CD133 human umbilical cord progenitor (stem) cells as part of a six month pilot study (2004). No bone marrow ablation or use of immu- nosuppressant drugs was involved. All the children showed clinically significant gains in mobility and cognitive function. Six children (75%) were re- ported to have improved in multiple areas of motor function by the end of the study [13]. One corti- cally blind, aphasic child began tracking objects visually and speaking using brief phrases.

In addition to these children, Ramirez and his associates have treated pediatric and adult patients suffering from a variety of physical challenges in- cluding macular degeneration, diabetic neuropathy, primary and secondary progressive multiple sclero- sis, traumatic brain injury, and emphysema. The Steenblock Research Institute (with which the au- thor is affiliated) provided technical support to this research-oriented clinical application of hUCSC in the form of data collection and analysis, patient interviews and other forms of follow-up, as well as development of protocols to enhance stem cell de- livery and post-treatment activity (USPTO Patent Pending). The results to-date has in most instances been very encouraging.

What is especially intriguing is that several cor- tically blind, aphasic, severely locomotor impaired children who received hUCSC in Mexico experi- enced amelioration of these during the first 120 days following treatment. Recently (May 2005) a female child with advanced genetic metachromatic leukodystrophy also experienced a resolution of cortical blindness and motor function gains in the first 60 days following a hUCSC injection. In addi- tion, at least 5 children with mild to moderate sei- zure activity prior to hUCSCT experienced a sig- nificant reduction in this and were able to reduce reliance on anti-seizure medications and even stop them in a few instances.

By far the best responder to the hUCSC being administered by Ramirez et al. has been cerebral palsy and TBI in children. The study cited above involved only eight children with CP, but a total of
13 with CP have been treated to date (June 2005). As indicated, all eight of those in the pilot study have shown clinically significant improvements in at least 7 areas of function (Caregiver assessment using a specially designed questionnaire one month prior to hUCSCT, then 1 month, 3 months and 6 months following the treatment. Questions were taken from McKeena’s report on side effects from cord blood and included changes in heart rate and blood pressure, nausea, back pain, rashes, chills, excessive thirst, rapid breathing, headaches, and others. There was also a section on symptoms and symptom improvements [14]. Eighty-five percent (85%) of the total complement of 13 children treated since March 2003 has experienced moderate or better improvement in their condition (2005 Caregiver survey). Four children and young people with TBIs were treated during 2004 with 75% re- ported to have experienced clinically significant improvements in overall cognitive and motor skill function.

This data is far from conclusive, though we at SRI feel it is compelling enough to justify addi- tional, more rigorous study (SRI has, in fact, begun the pre-IND process with the F.D.A. requisite to ultimately gaining approval to conduct a double blind, placebo controlled crossover trial involving use of hUCSC in children with cerebral palsy). These admitted limitations aside, if it is granted that some favorable neurologic influence is being exerted by these treatments, one is left with ques- tions concerning how these cells are bringing this about. If these cells were being given by IV, then a probable mechanism would be obvious: Some of the hUCSC are getting into the brain. The body of studies indicating these cells can be differentiated into neurons in the Petri dish [15-17] and penetrate the blood brain barrier [18-19] readily suggest that this would be the operative mechanism in cases in which hUCSC are infused by IV drip (something done by Ramirez et al for adult patients with neu- rologic diseases and insults). However, Dr. Rami- rez injected ~1.5 million hUCSCs subcutaneously near the umbilicus in all his pediatric patients. What mechanism would most likely account for the neurologic improvements seen following this? Es- pecially those that take place literally within the first day or so following treatment?


HYPOTHESIS

In reviewing the responses of children to the subcutaneously injected hUCSC during the first 72 hours or so following the treatment, I noticed that a few telltale side effects were cropping up: Many of the children experienced transient fluctuations in appetite in the first few weeks following the injec- tion, as well as mildly elevated body temperature (for a few days to a week or so), uncharacteristic sleepiness, and in some instances a mild rash and muscular fasciculations in the region adjacent to the injection site. These symptoms and subsequent medical testing did not indicate a graft-versus-host or host-versus-graft reaction was taking place.

While growth factors contained in the media in which the stem cells were suffused could account for some of the side effects ― most of which were abolished following total wash out of these growth factors ― some remained such as altered appetite, sleepiness and other signs that the injection was eliciting a response in the injection site (adipose tissue) that was influencing various neurologic cen- ters in the brain.

This body of evidence and the sequence of events noted readily suggest a hypothetical mecha- nism: Namely, that hUCSC injected in adipose tis- sue stimulates adipocytes to synthesize and express TNF-alpha, which in turn disrupts the blood brain barrier, as well as upregulates production of nerve growth factor (NGF). NGF produced by the adipo- cytes then enters the CNS where it exerts an influ- ence over cell growth, differentiation and function, and in addition affects cells of the immune system as well as inflammatory responses [20]. This adi- pocyte-generated NGF may also help quell seizures in children prone to them.

Normally, NGF crosses the blood brain barrier poorly if at all, but with permeability increased by TNF-alpha should gain access more readily and swiftly. It is the entry of NGF into the brain that makes explicable the many instances of neurologic improvement being reported as occurring during the first few hours following treatment. For while neurotrophins like NGF generally act slowly, it has been shown that this class of compounds also elic- its rapid signaling that modulates and influences a diverse range of cellular functions such as synaptic transmission, membrane excitability and activity- dependent synaptic plasticity. These rapid action effects are mediated primarily through the interac- tion of Trk receptors with ionotropic receptors and ion channels in the plasma membrane [21].

A review of the literature produced numerous laboratory and animal studies that lend credence to this hypothetical mechanism: Adipocytes exposed to proinflammatory stimuli such as lipopolysaccaride, or foreign antigens such as would be present in introduced hUCSC have been shown to begin synthesizing tumor necrosis factor-alpha (TNF-alpha) [22]. TNF-alpha, in turn, has been shown to induce white adipocytes to syn- thesize NGF [23] which belongs to a family of neu- rotrophins that have been shown to induce the sur- vival and proliferation of neurons. In addition, TNF-alpha has also demonstrated a blood brain barrier disrupting effect [24], and inhibited seizures in mice [25]. Interestingly, the latter effect parallels what we at SRI have seen take place in many chil- dren with a history of seizure activity, i.e., diminu- tion and in some cases total cessation of seizures during the first few weeks following hUCSCT.


SUMMATION AND CONCLUDING REMARKS

Since March 2003 sixteen children with cerebral palsy and TBI have been treated in Mexico (by Fernando Ramirez et al] using a subcutaneous in- jection of 1.5 million CD34+/CD133 human um- bilical cord stem cells in adipose tissue adjacent to the umbilicus. Eight-five percent of those with CP and 75% of those with TBI experienced clinically significant improvements in cognitive and motor skill function following this. What is intriguing is that many of these children began demonstrating benefit within the first day or so of receiving the injection. For example, many of the children began moving limbs that had previously were immobile, some grabbed objects with hands that previously were unable to uncurl and fasten around any object, and so forth. A few who demonstrated this kind and type of change were infants and toddlers.

Clearly too little time elapsed to attribute these positive changes to hUCSC migration to the brain, engraftment and proliferation.

However, these early onset clinically significant improvements become explicable when viewed as the end result of growth factor and neurotrophin activity. It is my contention (hypothesis) that the hUCSC deposited in adipose tissue causes adipo- cytes to synthesize blood brain barrier disruptive TNF-alpha and NGF. This would be consistent with published laboratory and animal studies, and with the rapid improvements seen in the treated children.

This hypothesis can be tested by doing quantita- tive assays of patients prior to hUCSCT and at regular intervals thereafter with respect to TNF- alpha and NGF. If the posited mechanism is at work, then a substantial increase in both TNF-alpha and NGF should be seen following injection of CD34+/CD133 human umbilical cord stem cells.

If this hypothesis is ultimately validated, the next logical step would be to mimic the effect of the human umbilical cord stem cells (hUCSC) by infusing patients with NGF and possibly the drug Mannitol to facilitate increasing the permeability of the blood brain barrier. Clinical responses obtained in these patients would then be compared and con- trasted to those seen in age, sex, and disease and disability matched patients who receive hUCSC only. The outcome would help determine if infused NGF alone can bring about the type and kind of neurologic improvement seen in the patients treated by Ramirez et al., or conversely point to benefits induced or facilitated by hUCSC that NGF alone cannot match or rival. In either case, progress in the treatment and amelioration of neurologic chal- lenges such as cerebral palsy in children would be at-hand.


REFERENCES

[1] WADHWA P, LAZARUS H, KOC ON, JAROSCA KJ, WOO D, STEVENS CE, RUBENSTEIN P AND ALUGHLIN MJ [2003] Hematopoietic recovery after unrelated umbilical cord- blood allogeneic transplantation in adults treated with in vivo stem cell factor (R-MetHusCF) and filgrastim ad- ministration. Leukemia Research 27: 215-220.
[2] Paquette R, Dergham S, Karpf E, Wang HJ, Salmon DJ, Souza L and Glazpy JA [2000] Ex vivo expanded unse- lected peripheral blood: progenitor cells reduce post- transplantation neutropenia, thrombocytopenia, and anemia in patients with breast cancer. Blood 96: 2385-
2390.
[3] MEAGHER R AND KLINGEMANN H [2002] Human Umbili- cal cord blood cells: How useful are they for the clini- cian? J Hemat Stem Cell Res 1: 445-448.
[4] CROOP J, COOPER R, FERNANDEX C, GRAVES V, KREISSMAN S, HANENBERG H, SMITH FO AND WILLIAMS DA [2003] Mobilization and collection of peripheral blood CD34+ cells from patients with Fanconi anemia. Blood 98: 2917-2921.
[5] GARBUZOVA-DAVIS S, WILLING AE, ZIGOVA T, SAPORTA S, JUSTEN EB, LANE JC, HUDSON JE, CHEN N, DAVIS CD AND SANBERG PR [2003] Intravenous administration of human umbilical cord blood cells in a mouse model of amyotrophic lateral sclerosis: Distribution, migration, and differentiation. J Hematother Stem Cell Res 12: 255-
270.
[6] ENDE N, CHEN R AND ENDE-HARRIS D [2001] Human umbilical cord blood cells ameliorate Alzheimer’s dis- ease in transgenic mice. J Med 32: 241-247.
[7] ENDE N AND CHEN R [2001] Human umbilical cord blood cells ameliorate Huntington’s disease in transgenic mice. J Med 32: 231-240.
[8] CHEN J, SANBERG P, LI Y, WANG L, LU M, WILLING AE, SANCHEZ-RAMOS J AND CHOPP M [2001] Intravenous administration of human umbilical cord blood reduces behavioral deficits after stroke in rats. Stroke 32: 2682-
2688.
[9] VENDRAME M, CASSADY J, NEWCOMB J, ET AL. [2004] Infusion of human umbilical cord blood cells in a rat model of stroke dose-dependently rescues behavioral deficits and reduces infarct volume. Stroke 35: 2390-
2395.
[10] KOGLER G, SENSKEN S, AIREY JA, TRAPP T, MUSCHEN M, FELDHAHN N, LIEDTKE S, SORG RV, FISCHER J, ROSENBAUM C, GRESCHAT S, KNIPPER A, BENDER J, DEGISTIRICI O, GAO J, CAPLAN AI, COLLETTI EJ, ALMEIDA-PORADA G, MULLER HW, ZANJANI E AND WERNET P [2004] A new human somatic stem cell from placental cord blood with intrinsic pluripotent differen- tiation potential. Exp Med 200: 123-135.
[11] IBID
[12] CHEN J, SANBERG PR, LI Y, WANG L, LU M, WILLING AE, SANCHEZ-RAMOS J AND CHOPP M [2001] Intravenous administration of human umbilical cord blood reduces behavioral deficits after stroke in rats. Stroke 32: 2682-
2688.
[13] STEENBLOCK DA, PAYNE AG AND DARNALL L [2005] Stem cells isolated from human umbilical cord blood for cerebral palsy. Manuscript submitted.
[14] MCKEENA DH, WAGNER JE AND MCCULLOUGH J [2003] Umbilical cord blood infusions are associated with mild reactions and are overall well tolerated. The 9th Annual Meeting of the International Society for Cellular Ther- apy, Phoenix, Arizona, May 29 - June 1, 2003.
[15] NEWMAN MB, DAVIS CD, KUZMIN-NICHOLS N AND SAN- BERG PR [2003] Human umbilical cord blood (HUCB) cells for central nervous system repair. Neurotox Res 5:
355-368.
[16] SANCHEZ-RAMOS JR [2002] Neural cells derived from adult bone marrow and umbilical cord blood. J Neurosci Res 69: 880-893.
[17] CHEN J, SANBERG PR, LI Y, WANG L, LU M, WILLING AE, SANCHEZ-RAMOS J AND CHOPP M [2001] Intravenous administration of human umbilical cord blood reduces behavioral deficits after stroke in rats. Stroke 32: 2682-
288.
[18] GARBUZOVA-DAVIS S, WILLING AE, ZIGOVA T, SAPORTA S, JUSTEN EB, LANE JC, HUDSON JE, CHEN N, DAVIS CD AND SANBERG PR [2003] Intravenous administration of human umbilical cord blood cells in a mouse model of amyotrophic lateral sclerosis: distribution, migration, and differentiation. J Hematother Stem Cell Res 12: 255-270.
[19] LU D, SANBERG PR, MAHMOOD A, LI Y, WANG L, SAN- CHEZ-RAMOS J AND CHOPP M [2002] Intravenous admini- stration of human umbilical cord blood reduces neuro- logical deficit in the rat after traumatic brain injury. Cell Transplant 11: 275-281.
[20] RITA LM. Analysis of the functional significance and mechanism of action of nerve growth factor’, Istituto di Neurobiologia-Roma, http://www.aging.cnr.it/uoe/uo1_107.htm.
[21] BLUM R AND KONNERTH A [2005] Neurotrophin- mediated rapid signaling in the central nervous system: mechanisms and functions. Physiology (Bethesda) 20:
70-78.
[22] COPPACK SW [2001] Pro-inflammatory cytokines and adipose tissue. Proc Ntri Soc 60: 349-356.
[23] PERAULLY, MR, JENKINS JR AND TAYHURN P [2004] NGF gene expression and secretion in white adipose tissue: regulation in 3T3-L1 adipocytes by hormones and in- flammatory cytokines. Amer J Physiol Endrocrinol Me- tab 2004 287: E331-E339.
[24] MAYHAN WG [2002] Cellular mechanisms by which tumor necrosis factor-alpha produces disruption of the blood-brain barrier. Brain Res 927: 144-152.
[25] BALOSSO S, RAVIZZA T, PEREGO C, PESHON J, CAMPBELL IL, DE SIMONI MG AND VEZZANI A [2005] Tumor necro- sis factor-alpha inhibits seizures in mice via p75 recep- tors. Ann Neurol 57: 804-812.
0 Comments
Posted on 2013 Oct 25 by admin
Name:
E-mail: (optional)
Smile:

Captcha
Powered by CuteNews