Vitamin D History

1645 Whistler (1650 Glisson): rickets described 1920 Mellanby: dogs raised indoors (no sunlight) developed rickets: cod liver oil cured it 1920s McCollum: bubbling oxygen through a preparation of fat-soluble vitamins inactivated vitamin A but not vitamin D 1923 Goldblatt and Soames: skin produced a substance equivalent to vitamin D when irradiated by sunlight or UV light 1920s Hess and Weinstock: skin irradiated with UV light and fed to rats cured rickets

Rickets
Mineralization defect Hypertrophy of Chondrocytes (cartilage)

Short stature
Bony deformities In children: bowed legs defects in rib cage

Vitamin D3 = Cholecalciferol Produced in animals Vitamin D2 = Ergocalciferol Derived from a precursor found in plants and yeast

Dietary Sources of Vitamin D

Fatty fish (mackerel and salmon) Fish oils (cod and tuna liver oils) Foods fortified with Vitamin D: Milk Cereals Breads Fortified foods rarely contain the labeled amount of vitamin D

Vitamin D is a fat soluble vitamin

Hydrophobic Poorly soluble in aqueous environments such as the intestinal lumen, plasma, and the cytoplasm of cells Hence, requires carriers or transport proteins in aqueous environments Easily soluble in lipid-rich environments so, crosses membranes readily and is stored in lipid droplets within cells

Digestion and Absorption:

Fat soluble vitamin: emulsified with lipids and bile acids

Taken up into enterocytes by passive diffusion

Vitamin D is packaged with dietary lipids into chylomicrons, secreted into lymph, then circulates in plasma. Action of lipoprotein lipase removes lipids into muscle and fat; some vitamin D stored in fat, as well. Chylomicron remnants then taken up by liver.

7-Dehydrocholesterol can be converted to Vitamin D in the skin; requires UV light

Vitamin D not strictly a vitamin since it can be produced by the body

The amount of Vitamin D produced in the skin is reduced with the use of sunscreen
1 minimal erythemal dose (minimum sunburn) used: equivalent to 10,000-25,000 IU of oral vitamin D (single exposure) SPF 8 sunscreen used in this example Clothing also reduces sun exposure

The ability to produce vitamin D in the skin varies with:

Pigmentation: melanin levels reduce UVB to skin Age:
Young 20-30 Elderly 62-80 1 exposure of minimal erythemal dose

70 yrs: 75% reduction in vit.D production in skin (reduced 7-dehydrocholesterol in the skin)

Vitamin D production in skin varies with:

Time of Day

Season of year
Latitude: 42 = Boston; sunlight too dim to produce vitamin D in the skin from November through February Stores of vitamin D in fat may be adequate to provide vitamin D in the winter 3 weekly exposures of hands and face for 20 min

Vitamin D is converted to 25-hydroxyvitamin D3 in the liver (and also skin, intestine, and kidney) 25-OH vit D3 is exported from liver on vitamin D-binding protein (DBP) Metabolism to 1,25-dihydroxyvitamin D3 occurs in kidney; this is the active form 24,25(OH)2 vit. D3 is much less active

Metabolism of Vitamin D:
Hydroxylation of carbon 25 to make 25-OH vit. D3 by 25-hydroxylase occurs primarily in liver (also skin, intestine and kidney) Export of 25-OH vit. D3 from liver into circulation on DBP

25-hydroxylation is poorly regulated, hence, 25-hydroxyvitamin D levels in plasma are used to determine vitamin D status Levels of 25-OH most accurately reflect dietary intake and cutaneous production, since vit. D3 is rapidly converted to 25-OH vit. D3 and this is the major circulating form

More on Metabolism
Metabolism of 25-hydroxyvitamin D3 to the biologically active form occurs in the kidney: hydroxylation of carbon 1 to make 1,25-dihydroxyvitamin D3: export from kidney on DBP Other tissues have the 1-hydroxylase, but contribute little to 1,25-(OH)2D levels Placenta during pregnancy Macrophages, other cells 24-hydroxylase in kidney: makes 24,25-dihydroxyvitamin D: less active, unknown function, step in degradation Excreted form: calcitroic acid

Functions of 1,25 (OH)2Vitamin D3

Functions as a hormone

Primary function in whole body calcium and phosphorous homeostasis

Maintenance of a healthy skeleton Maintain serum calcium (10 mg/ 100 ml) and phosphorous levels (4 mg/100 ml) for bone mineralization

1,25 (OH)2Vit.D3 increases calcium levels:

3 mechanisms: Most important: increases intestinal absorption of calcium Increases renal reabsorption of calcium Increases calcium mobilization from bone

1,25(OH)2Vit. D3 is a hormone that binds to the Vitamin D Receptor, a transcription factor

DNA binding domain: 2 zinc fingers Ligand binding domain: binds 1,25(OH)2vit. D3 Heterodimerizes with RXR (retinoid X receptor) Binds to VDREs in the promoters of many genes

Hormonal action of 1,25(OH)2vitamin D3

Both increases and decreases in transcription of genes

Induction

CaT1=TRPV6

9-cis retinoic acid

Repression of transcription

1,25(OH)2Vit.D3 action on Intestine

Increases synthesis of proteins involved in the uptake and transport of calcium Increases transcription of genes: TRPV6/CaT1 calcium channel (epithelium) Calbindin, a cellular calcium binding protein (Ca2+ movement in cytoplasm) Basolateral calcium pump (export) Directly increases calcium absorption across enterocyte plasma membrane

Increases phosphorous absorption in small intestine by increasing expression of Na-Pi co-transporter called Npt2

1,25(OH)2Vit.D3 Actions on Bone

Bone is continually remodeled In osteoblasts, 1,25 (OH)2D3 increases the expression of osteopontin and osteocalcin, bone matrix proteins Promotes mineralization by maintenance of serum calcium and phosphorous levels When dietary calcium is low, 1,25(OH)2D3 promotes osteoclast differentiation (involved in bone resorption; for mobilization of bone calcium)

1,25(OH)2Vit.D3 Action on Kidney

1,25(OH)2D3 suppresses 1-hydroxylase activity (decreases further production of 1,25(OH)2D3) Stimulates 24-hydroxylase activity: step towards removal of excess 1,25(OH)2D3 from body Enhances renal calcium reabsorption

1,25(OH)2Vit.D3 regulates genes unrelated to calcium homeostasis

Alters transcription of genes involved in the cell cycle to inhibit proliferation and induce terminal differentiation c-myc, c-fos, c-cis (oncogenes) Used to treat psoriasis, a hyperproliferative skin disorder Stimulates immune function Modulates muscle cell calcium levels and cell growth Pancreas: enhances insulin secretion Role in female fertility Nervous system: antiproliferative, prodifferentiation

Regulation of Vitamin D Metabolism

25-hydroxylase activity in liver is not well regulated 1-hydroxylase activity in kidney is highly regulated through transcription Low 1,25(OH)2D3 in circulation: increase 1-hydroxylase activity to increase 1,25(OH)2D3 production High 1,25(OH)2D3 levels: decrease 1-hydroxylase activity and increase 24-hydroxylase activity

More on Regulation of Vitamin D

With vitamin D deficiency, intestinal calcium absorption decreases from 30-50% to 10-15%; circulating calcium decreases Parathyroid gland calcium sensor detects Ca2+ levels in serum and increases synthesis and production of parathyroid hormone (PTH) PTH increases renal 1-hydroxylase activity, to increase production of 1,25(OH)2Vit D3, increases renal reabsorption of calcium, mobilizes osteoclasts to mobilize bone calcium (works through a signaling pathway on pre-osteoblasts)

Summary of Vitamin D regulatory Pathway: High PTH

25(OH)D3
1-OHase

Low Pi Low Calcium 1,25(OH) D 2 3

24- OHase

High calcium Low PTH High Pi High 1,25(OH)2D3

24,25(OH)2D3

VDR is essential for post-natal development

Knockout mouse: targeted disruption of VDR gene Animals normal at birth, retarded growth after birth, then rickets Developed alopecia by 7 weeks Males and females infertile Knockout 1-OHase: rickets, females infertile Can somewhat cure with high Ca2+, Pi Knockout mouse: 24-OHase Defective mineralization of bone

Non-VDRnuc mediated effects of Vitamin D

Mediated by a vitamin D receptor on the plasma membrane called VDRmem VDRmem is not related to VDRnuc Prefers 1, 25 (OH)2 Vit D3 in the cis form Initiates intracellular signaling cascade involving PKC, phospholipase C, MAPK

Increases Calcium absorption by rapid opening of Ca2+ and Cl- channels

Summary Low circulating calcium increases PTH secretion High calcium inhibits PTH secretion High 1,25(OH)2vit. D3 inhibits PTH secretion Phosphate regulates PTH, but less effectively; increased P042increases PTH indirectly

Vitamin D Deficiency
Serum levels of 25(OH)Vit D3 <10 g/L Rickets: newly synthesized bones not properly mineralized: soft bones Short stature and bony deformities Can also be caused by genetic block in 1-hydroxylase, or mutant VDR Osteomalacia in adults: poor mineralization, decreased opacity of bones, increased fracture risk

Dietary Recommendations (2010)

RDAs established for first time (assuming all Vit D obtained from diet) Adequate Intakes for infants (1 g = 40 IU) g vit. D/day Infants (birth-1 y) (AI) 10 Children and adults <51 y 15 Adults 51-70 y 15 Adults 71+ y 20 Pregnant and lactating women 15
12 oz. of fortified whole milk; 12 oz. of fortified orange juice; Small portions of fish (100 g usually has RDA)

Vitamin D Toxicity
Results from >10,000 IU/day for months Cannot get this easily from dietary sources Unlikely from exposure to sunlight Hypercalcemia Hyperphosphatemia Hypertension Anorexia Nausea Renal failure Death

Excess exposure to sunlight can produce large doses of Vitamin D3 in skin Unlikely to result in toxicity due to further metabolism of Vit. D in skin exposed to sunlight lumisterol tachysterol suprasterol I & II Inactive forms of Vitamin D

Tolerable upper intake levels of Vitamin D (2010):

Infants 0-1 y Children 4-8 y 62.5 g/day (2500 IU) 75 g/day (3000 IU)

All others (8 y up)

100 g/day (4000 IU)

Vitamin D intoxication >150 g/L of 25(OH)Vit D3 in serum Vitamin D deficiency <11 g/L

Decreased calcium absorption due to decreased calcium intake or decreased vitamin D leads to an increased risk of fractures

Factors preserving BMD

Calcium and VitaminD supplementation of diet

Exercise
Adequate dietary protein levels Hormone replacement therapy for post-menopausal women bone turnover rate by 10-15% BMD by 2-5% fracture incidence by 25% Adequate Vitamin K intake